Yellow Field Pea Cultivar 3997499

ABSTRACT

A field pea cultivar designated 3997499 is disclosed. Embodiments include the seeds, plants, and plant parts of field pea cultivar 3997499, and methods for producing a field pea plants by crossing field pea cultivar 3997499 with itself or with another field pea variety. Embodiments further include methods for producing field pea plants containing one or more genes or transgenes and the transgenic field pea plants and plant parts produced by those methods. Embodiments further relate to field pea cultivars, breeding cultivars, plant parts, and cells derived from field pea cultivar 3997499, methods for producing other field pea cultivars, lines, hybrids, or plant parts derived from field pea cultivar 3997499, and the field pea plants, varieties, and their parts derived from use of those methods.

BACKGROUND

All publications cited in this application are herein incorporated byreference.

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possesses the traits tomeet the program goals. The goal is to combine in a single variety animproved combination of desirable traits from the parental germplasm.These important traits may include higher seed yield, resistance todiseases and insects, better stems and roots, tolerance to drought andheat, and better agronomic quality.

Yellow field pea, Pisum sativum, is an important and valuable fieldcrop. Thus, a continuing goal of pea plant breeders is to developstable, high yielding pea cultivars that are agronomically sound. Thereasons for this goal are to maximize the amount of grain produced onthe land used and to supply food for both animals and humans. Toaccomplish this goal, the pea breeder must select and develop pea plantsthat have traits that result in superior cultivars.

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

SUMMARY

It is to be understood that the embodiments include a variety ofdifferent versions or embodiments, and this Summary is not meant to belimiting or all-inclusive. This Summary provides some generaldescriptions of some of the embodiments, but may also include some morespecific descriptions of other embodiments.

An embodiment provides a field pea cultivar designated 3997499. Anotherembodiment relates to the seeds of field pea cultivar 3997499, to theplants of field pea cultivar 3997499 and to methods for producing a peaplant produced by crossing field pea cultivar 3997499 with itself oranother field pea cultivar, and the creation of variants by mutagenesisor transformation of field pea cultivar 3997499.

Any such methods using the field pea cultivar 3997499 are a furtherembodiment: selfing, backcrosses, hybrid production, haploid production,crosses to populations, and the like. All plants produced using fieldpea cultivar 3997499 as at least one parent are within the scope of theembodiments. Advantageously, field pea cultivar 3997499 could be used incrosses with other, different pea plants to produce first generation(Fi) pea hybrid seeds and plants with superior characteristics.

Another embodiment provides for single or multiple gene converted plantsof field pea cultivar 3997499. The transferred gene(s) may be a dominantor recessive allele. The transferred gene(s) may confer such traits asherbicide resistance, insect resistance, resistance for bacterial,fungal, or viral disease, male fertility, male sterility, enhancednutritional quality, modified fatty acid metabolism, modifiedcarbohydrate metabolism, modified seed yield, modified oil percent,modified protein percent, modified shattering, modified iron-deficiencychlorosis, and industrial usage. The gene may be a naturally occurringpea gene or a transgene introduced through genetic engineeringtechniques. Another embodiment provides methods for using field peacultivar 3997499 as source material for producing haploid or apomicticfield pea plants.

Another embodiment provides for regenerable cells for use in tissueculture of field pea cultivar 3997499. The tissue culture may be capableof regenerating plants having all the physiological and morphologicalcharacteristics of the foregoing pea plant, and of regenerating plantshaving substantially the same genotype as the foregoing pea plant. Theregenerable cells in such tissue cultures may be embryos, protoplasts,meristematic cells, callus, pollen, leaves, ovules, anthers, cotyledons,hypocotyl, pistils, roots, root tips, flowers, seeds, petiole, stipules,vine, tendril, pods, or stems. Still a further embodiment provides forpea plants regenerated from the tissue cultures of field pea cultivar3997499.

Another embodiment provides for a method of editing the genome of fieldpea cultivar plant 3997499, said method comprising editing the genome ofthe plant, or plant part thereof, of field pea cultivar 3997499, whereinsaid method is selected from the group comprising zinc finger nucleases,transcription activator-like effector nucleases (TALENs), engineeredhoming endonucleases/meganucleases, and the clustered regularlyinterspaced short palindromic repeat (CRISPR)-associated protein9 (Cas9)system.

The pea seed of field pea cultivar 3997499 may be provided as anessentially homogeneous population of field pea cultivar 3997499.Essentially homogeneous populations of seed are generally free fromsubstantial numbers of other seed.

As used herein, “at least one,” “one or more,” and “and/or” areopen-ended expressions that are both conjunctive and disjunctive inoperation. For example, each of the expressions “at least one of A, Band C,” “at least one of A, B, or C,” “one or more of A, B, and C,” “oneor more of A, B, or C” and “A, B, and/or C” means A alone, B alone, Calone, A and B together, A and C together, B and C together, or A, B andC together.

As used herein, “sometime” means at some indefinite or indeterminatepoint of time. So for example, as used herein, “sometime after” meansfollowing, whether immediately following or at some indefinite orindeterminate point of time following the prior act.

Various embodiments are set forth in the Detailed Description asprovided herein and as embodied by the claims. It should be understood,however, that this Summary does not contain all of the aspects andembodiments, is not meant to be limiting or restrictive in any manner,and that embodiment(s) as disclosed herein is/are understood by those ofordinary skill in the art to encompass obvious improvements andmodifications thereto.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by study of thefollowing descriptions.

DEFINITIONS

In the description and tables herein, a number of terms are used. Inorder to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

Cotyledon. A cotyledon is a type of seed leaf. The cotyledon containsthe food storage tissues of the seed.

Embryo. The embryo is the small plant contained within a mature seed.

F₃. The “F3” symbol denotes a generation resulting from the selfing ofthe F2 generation along with selection for type and rogueing ofoff-types. The “F” number is a term commonly used in genetics, anddesignates the number of the filial generation. The “F₃” generationdenotes the offspring resulting from the selfing or self-mating ofmembers of the generation having the next lower “F” number, that is, the“F₂” generation.

Gene. Gene refers to a segment of nucleic acid. A gene can be introducedinto a genome of a species, whether from a different species or from thesame species, using transformation, gene editing techniques, or variousbreeding methods.

Hilum. Hilum refers to the scar left on the seed that marks the placewhere the seed was attached to the pod prior to the seed beingharvested.

Hypocotyl. A hypocotyl is the portion of an embryo or seedling betweenthe cotyledons and the root. Therefore, it can be considered atransition zone between shoot and root.

Iron Deficiency Chlorosis. Iron deficiency chlorosis (IDC) is ayellowing of the leaves caused by a lack of iron in the pea plant. Ironis essential in the formation of chlorophyll, which gives plants theirgreen color. In high pH soils iron becomes insoluble and cannot beabsorbed by plant roots. Pea cultivars differ in their genetic abilityto utilize the available iron.

Linoleic Acid Percent. Linoleic acid is one of the most abundant fattyacids in field pea seeds and can be measured by gas chromatography.

Locus. A locus confers one or more traits such as, for example, malesterility, herbicide tolerance, insect resistance, disease resistance,waxy starch, modified fatty acid metabolism, modified phytic acidmetabolism, modified carbohydrate metabolism, and modified proteinmetabolism. The trait may be, for example, conferred by a naturallyoccurring gene introduced into the genome of the variety by geneediting, backcrossing, a natural or induced mutation, or a transgeneintroduced through genetic transformation techniques. A locus maycomprise one or more alleles integrated at a single chromosomallocation.

Oil or Oil Percent. Pea seeds contain a considerable amount of oil. Oilis reported as a percentage basis.

Oleic Acid Percent. Oleic acid is one of the most abundant fatty acidsin field pea seeds and is measured by gas chromatography and is reportedas a percent of the total oil content.

Palmitic Acid Percent. Palmitic acid is one of the most abundant fattyacids in field pea seeds and is measured by gas chromatography and isreported as a percent of the total oil content.

Plant. A plant refers to a whole plant, any part thereof, or a cell ortissue culture derived from a plant, comprising any of: whole plants,plant components or organs (e.g., leaves, stems, roots, etc.), planttissues, seeds, embryos, plant cells, protoplasts and/or progeny of thesame. A plant cell is a biological cell of a plant, taken from a plantor derived through culture of a cell taken from a plant.

Plant Height. Plant height is taken from the top of the soil to the topnode of the plant and is measured in centimeters.

Plant Parts. Plant parts (or a pea plant, or a part thereof) includesbut is not limited to protoplasts, cells, leaves, stems, roots, roottips, anthers, pistils, seed, grain, embryo, pollen, ovules, cotyledon,hypocotyl, pod, flower, shoot, tissue, petiole, cells, meristematiccells, stipules, vine, tendril, and the like.

Pod. It consists of the shell or wall (pericarp) and the pea seeds.

Progeny. Progeny includes an Fi pea plant produced from the cross of twofield pea plants where at least one plant includes field pea cultivar3997499 and progeny further includes, but is not limited to, subsequentF₂, F₃, F₄, F₅, F₆, F₇, F₈, F₉, and F₁₀ generational crosses with therecurrent parental line.

Protein Percent. Pea seeds contain a considerable amount of protein.Protein is generally measured by NIR spectrophotometry, but can bemeasured by other means well-known in the art.

Pubescence. Pubescence refers to a covering of very fine hairs closelyarranged on the leaves, stems, vines, and pods of the pea plant.

Maturity. Early to medium maturity is defined as being around 75 to 85days from planting in an ideal crop growth testing environment.

Grams Per 1000 Seeds. Pea seeds vary in seed size; therefore, the numberof seeds in 1000 grams also varies.

Single Gene Converted (Conversion). Single gene converted (conversion)plants refers to plants which are developed by a plant breedingtechnique called backcrossing wherein essentially all of the desiredmorphological and physiological characteristics of a variety arerecovered in addition to the single gene transferred into the varietyvia the backcrossing technique or via genetic engineering. Byessentially all of the morphological and physiological characteristics,it is meant that the characteristics of a plant are recovered that areotherwise present when compared in the same environment, other than anoccasional variant trait that might arise during backcrossing or directintroduction of a transgene.

Sulfonylurea Reaction. Sulfonylurea reaction refers to a plant’stolerance, resistance or susceptibility to sulfonylurea herbicides andrefers to a plant which contains the ALS gene, which confers resistanceto some of the sulfonylurea herbicides.

Trypsin. Trypsin is a digestive enzyme, specifically, a pancreaticserine protease enzyme with substrate specificity based upon positivelycharged lysine and arginine side chains and is excreted by the pancreas.Trypsin aids in the digestion of food proteins and other biologicalprocesses.

Trypsin inhibitor units. Trypsin inhibitor units or abbreviated as TIU,is an assay measuring the quantity of trypsin inhibitor in a field peaseed or field pea product thereof. Measurement of trypsin inhibitorunits is a technique well-known in the art.

DETAILED DESCRIPTION

Yellow field pea cultivar 3997499 has a medium maturity and is resistantto Powdery Mildew.

Yellow field pea cultivar 3997499 has shown uniformity and stability, asdescribed in the following variety description information. Yellow fieldpea cultivar 3997499 has been self-pollinated a sufficient number ofgenerations with careful attention to uniformity of plant type and hasbeen increased with continued observation for uniformity.

Yellow field pea cultivar 3997499 has the following morphologic andother characteristics based primarily on field data collected in NorthDakota and Manitoba, Canada during 2020, 2021 and 2022.

TABLE 1 VARIETY DESCRIPTION INFORMATION Characteristic 3997499 Planttype Yellow Field Pea Plant habit Determinate Plant height Greater than50 cm Stem fasciation Absent Presence of leaflets Semi-leafless MaturityMedium Flower color White Stipules Semi-leafless Time of floweringMedium Protein content on dry basis 28.4% Stipule development Normal Podlength (observed at first flowering node) Medium Pod width (observed atfirst flowering node) Medium Pod color (immature) Light-green Podcurvature (fully swollen) Absent Seed shape Slightly round Weight(g/1000 seeds) 230 Color of cotyledon Yellow Black of hilum AbsentReaction to Powdery Mildew Resistant Seed size Medium - large Seedsurface Smooth Color pattern Monocolor Hilum White Primary colorYellow-green

Breeding With Yellow Field Pea Cultivar 3997499

The complexity of inheritance influences choice of the breeding method.Backcross breeding is used to transfer one or a few favorable genes fora highly heritable trait into a desirable cultivar. This approach hasbeen used extensively for breeding disease-resistant cultivars. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes. The use of recurrentselection in self-pollinating crops depends on the ease of pollination,the frequency of successful hybrids from each pollination, and thenumber of hybrid offspring from each successful cross.

Promising advanced breeding lines are thoroughly tested and compared toappropriate standards in environments representative of the commercialtarget area(s) for three or more years. The best lines are candidatesfor new commercial cultivars; those still deficient in a few traits maybe used as parents to produce new populations for further selection.

A population means a set comprising any number, including one, ofindividuals, objects, or data from which samples are taken forevaluation, e.g., estimating QTL effects and/or disease tolerance. Mostcommonly, the terms relate to a breeding population of plants from whichmembers are selected and crossed to produce progeny in a breedingprogram. A population of plants can include the progeny of a singlebreeding cross or a plurality of breeding crosses and can be eitheractual plants or plant derived material, or in silico representations ofplants. The member of a population need not be identical to thepopulation members selected for use in subsequent cycles of analyses nordoes it need to be identical to those population members ultimatelyselected to obtain a final progeny of plants. Often, a plant populationis derived from a single biparental cross but can also derive from twoor more crosses between the same or different parents. Although apopulation of plants can comprise any number of individuals, those ofskill in the art will recognize that plant breeders commonly usepopulation sizes ranging from one or two hundred individuals to severalthousand, and that the highest performing 5% to 20% of a population iswhat is commonly selected to be used in subsequent crosses in order toimprove the performance of subsequent generations of the population in aplant breeding program.

These processes, which lead to the final step of marketing anddistribution, usually take from eight to twelve years from the time thefirst cross is made. Therefore, development of new cultivars is atime-consuming process that requires precise forward planning, efficientuse of resources, and a minimum of changes in direction.

A most difficult task is the identification of individuals that aregenetically superior, because for most traits the true genotypic valueis masked by other confounding plant traits or environmental factors.One method of identifying a superior plant is to observe its performancerelative to other experimental plants and to a widely grown standardcultivar. If a single observation is inconclusive, replicatedobservations provide a better estimate of its genetic worth.

The goal of field pea plant breeding is to develop new and superiorfield pea cultivars and hybrids. The breeder initially selects andcrosses two or more parental lines, followed by repeated selfing andselection, producing many new genetic combinations. The breeder cantheoretically generate billions of different genetic combinations viacrossing, selection, selfing and mutations. Therefore, a breeder willnever develop the same line, or even very similar lines, having the samefield pea traits from the exact same parents.

Each year, the plant breeder selects the germplasm to advance to thenext generation. This germplasm is grown under different geographicalclimate and soil conditions and further selections are then made duringand at the end of the growing season. The cultivars that are developedare unpredictable because the breeder’s selection occurs in environmentswith no control at the DNA level, and with millions of differentpossible genetic combinations being generated. A breeder of ordinaryskill in the art cannot predict the final resulting lines he develops,except possibly in a very gross and general fashion. The same breedercannot produce the same cultivar twice by using the same originalparents and the same selection techniques. This unpredictability resultsin the expenditure of large amounts of research monies to developsuperior new field pea cultivars.

The development of new field pea cultivars requires the development andselection of field pea varieties, the crossing of these varieties andselection of superior hybrid crosses. The hybrid seed is produced bymanual crosses between selected male-fertile parents or by using malesterility systems. These hybrids are selected for certain single genetraits such as color, flower color, pubescence color or herbicideresistance which indicate that the seed is truly a hybrid. Additionaldata on parental lines, as well as the phenotype of the hybrid,influence the breeder’s decision whether to continue with the specifichybrid cross.

Breeding programs combine desirable traits from two or more cultivars orvarious broad-based sources into breeding pools from which cultivars aredeveloped by selfing and selection of desired phenotypes. Pedigreebreeding is used commonly for the improvement of self-pollinating crops.Two parents that possess favorable, complementary traits are crossed toproduce an F₁. An F₂ population is produced by selfing one or severalFis. Selection of the best individuals may begin in the F₂ population;then, beginning in the F₃, the best individuals in the best families areselected. Replicated testing of families can begin in the F₄ generationto improve the effectiveness of selection for traits with lowheritability. At an advanced stage of inbreeding (i.e., F₆ and F₇), thebest lines or mixtures of phenotypically similar lines are tested forpotential release as new cultivars.

As used herein, “fertilization” and/or “crossing” broadly includesbringing the genomes of gametes together to form zygotes but alsobroadly may include pollination, syngamy, fecundation and otherprocesses related to sexual reproduction. Typically, a cross and/orfertilization occurs after pollen is transferred from one flower toanother, but those of ordinary skill in the art will understand thatplant breeders can leverage their understanding of fertilization and theoverlapping steps of crossing, pollination, syngamy, and fecundation tocircumvent certain steps of the plant life cycle and yet achieveequivalent outcomes, for example, a plant or cell of a field peacultivar described herein. In certain embodiments, a user of thisinnovation can generate a plant of the claimed invention by removing agenome from its host gamete cell before syngamy and inserting it intothe nucleus of another cell. While this variation avoids the unnecessarysteps of pollination and syngamy and produces a cell that may notsatisfy certain definitions of a zygote, the process falls within thedefinition of fertilization and/or crossing as used herein whenperformed in conjunction with these teachings. In certain embodiments,the gametes are not different cell types (i.e., egg vs. sperm), butrather the same type and techniques are used to effect the combinationof their genomes into a regenerable cell. Other embodiments offertilization and/or crossing include circumstances where the gametesoriginate from the same parent plant, i.e., a “self' or“self-fertilization”. While selfing a plant does not require thetransfer pollen from one plant to another, those of skill in the artwill recognize that it nevertheless serves as an example of a cross,just as it serves as a type of fertilization. Thus, methods andcompositions taught herein are not limited to certain techniques orsteps that must be performed to create a plant or an offspring plant ofthe claimed invention, but rather include broadly any method that issubstantially the same and/or results in compositions of the claimedinvention. Crop Performance

Crop performance is used synonymously with plant performance and refersto of how well a plant grows under a set of environmental conditions andcultivation practices. Crop performance can be measured by any metric auser associates with a crop’s productivity (e.g., yield), appearanceand/or robustness (e.g., color, morphology, height, biomass, maturationrate), product quality (e.g., seed protein content, seed oil content,seed carbohydrate content, etc.), cost of goods sold (e.g., the cost ofcreating a seed, plant, or plant product in a commercial, research, orindustrial setting) and/or a plant’s tolerance to disease (e.g., aresponse associated with deliberate or spontaneous infection by apathogen), pests, microbes, fungi, and/or environmental stress (e.g.,drought, flooding, low nitrogen or other soil nutrients, wind, hail,temperature, day length, etc.). Crop performance can also be measured bydetermining a crop’s commercial value and/or by determining thelikelihood that a particular inbred, hybrid, or variety will become acommercial product, and/or by determining the likelihood that theoffspring of an inbred, hybrid, or variety will become a commercialproduct. Crop performance can be a quantity (e.g., the volume or weightof seed or other plant product measured in liters or grams) or someother metric assigned to some aspect of a plant that can be representedon a scale (i.e., assigning a 1 to10 value to a plant based on itsdisease tolerance).

A microbe will be understood to be a microorganism, i.e., a microscopicorganism, which can be single celled or multicellular. Microorganismsare very diverse and include all the bacteria, archaea, protozoa, fungi,and algae, especially cells of plant pathogens and/or plant symbionts.Certain animals are also considered microbes, e.g., rotifers. In variousembodiments, a microbe can be any of several different microscopicstages of a plant or animal. Microbes also include viruses, viroids, andprions, especially those which are pathogens or symbionts to cropplants.

A fungus includes any cell or tissue derived from a fungus, for examplewhole fungus, fungus components, organs, spores, hyphae, mycelium,and/or progeny of the same. A fungus cell is a biological cell of afungus, taken from a fungus or derived through culture of a cell takenfrom a fungus.

A pest is any organism that can affect the performance of a plant in anundesirable way. Common pests include microbes, animals (e.g., insectsand other herbivores), and/or plants (e.g., weeds). Thus, a pesticide isany substance that reduces the survivability and/or reproduction of apest, e.g., fungicides, bactericides, insecticides, herbicides, andother toxins.

Tolerance or improved tolerance in a plant to disease conditions (e.g.,growing in the presence of a pest) will be understood to mean anindication that the plant is less affected by the presence of pestsand/or disease conditions with respect to yield, survivability and/orother relevant agronomic measures, compared to a less tolerant, more“susceptible” plant. Tolerance is a relative term, indicating that a“tolerant” plant survives and/or performs better in the presence ofpests and/or disease conditions compared to other (less tolerant) plants(e.g., a different field pea cultivar) grown in similar circumstances.As used in the art, tolerance is sometimes used interchangeably with“resistance”, although resistance is sometimes used to indicate that aplant appears maximally tolerant to, or unaffected by, the presence ofdisease conditions. Plant breeders of ordinary skill in the art willappreciate that plant tolerance levels vary widely, often representing aspectrum of more-tolerant or less-tolerant phenotypes, and are thustrained to determine the relative tolerance of different plants, plantlines or plant families and recognize the phenotypic gradations oftolerance.

Desired Trait or Traits

In certain embodiments, plants disclosed herein can be modified toexhibit at least one desired trait, and/or combinations thereof. Theembodiments disclosed herein, are not limited to any set of traits thatcan be considered desirable, but nonlimiting examples include malesterility, herbicide tolerance, pest tolerance, disease tolerance,modified fatty acid metabolism, modified carbohydrate metabolism,modified seed yield, modified seed oil, modified seed protein, modifiedlodging resistance, modified shattering, modified iron-deficiencychlorosis, modified water use efficiency, and/or combinations thereof.Desired traits can also include traits that are deleterious to plantperformance, for example, when a researcher desires that a plantexhibits such a trait in order to study its effects on plantperformance.

Methods disclosed herein include conferring desired traits to plants,for example, by mutating sequences of a plant, introducing nucleic acidsinto plants, using plant breeding techniques and various crossingschemes, etc. These methods are not limited as to certain mechanisms ofhow the plant exhibits and/or expresses the desired trait. In certainnonlimiting embodiments, the trait is conferred to the plant byintroducing a nucleotide sequence (e.g., using plant transformationmethods) that encodes production of a certain protein by the plant. Incertain nonlimiting embodiments, the desired trait is conferred to aplant by causing a null mutation in the plant’s genome (e.g., when thedesired trait is reduced expression or no expression of a certaintrait). In certain nonlimiting embodiments, the desired trait isconferred to a plant by crossing two plants to create offspring thatexpress the desired trait. It is expected that users of these teachingswill employ a broad range of techniques and mechanisms known to bringabout the expression of a desired trait in a plant. Thus, as usedherein, conferring a desired trait to a plant is meant to include anyprocess that causes a plant to exhibit a desired trait, regardless ofthe specific techniques employed.

Using Yellow Field Pea Cultivar 3997499 to Develop Other Field PeaVarieties

Field pea varieties such as yellow field pea cultivar 3997499 aretypically developed for use in seed production. However, field peavarieties such as field pea cultivar 3997499 also provide a source ofbreeding material that may be used to develop new field pea varieties.Plant breeding techniques known in the art and used in a field pea plantbreeding program include, but are not limited to, recurrent selection,mass selection, bulk selection, mass selection, backcrossing, pedigreebreeding, open pollination breeding, restriction fragment lengthpolymorphism enhanced selection, speed breeding, genetic marker enhancedselection, making double haploids, and transformation. Oftencombinations of these techniques are used. The development of field peavarieties in a plant breeding program requires, in general, thedevelopment and evaluation of homozygous varieties. There are manyanalytical methods available to evaluate a new variety. The oldest andmost traditional method of analysis is the observation of phenotypictraits, but genotypic analysis may also be used.

Additional Breeding Methods

One embodiment is directed to methods for producing a field pea plant bycrossing a first parent field pea plant with a second parent field peaplant, wherein the first or second field pea plant is the field peaplant from field pea cultivar 3997499. Further, both first and secondparent field pea plants may be from either field pea cultivar 3997499.Therefore, any methods using field pea cultivar 3997499 are part of theembodiments: selfing, backcrosses, hybrid breeding, and crosses topopulations. Any plants produced using field pea cultivar 3997499 as atleast one parent are also within the scope of the embodiments. Any suchmethods using field pea variety 3997499 are part of the embodiments:selfing, sibbing, backcrosses, mass selection, pedigree breeding, bulkselection, hybrid production, crosses to populations, and the like.These methods are well known in the art and some of the more commonlyused breeding methods are described herein. Descriptions of breedingmethods can be found in one of several reference books (e.g., Allard,Principles of Plant Breeding (1960); Simmonds, Principles of CropImprovement (1979)).

The following describes breeding methods that may be used with field peacultivar 3997499 in the development of further field pea plants. Onesuch embodiment is a method for developing cultivar 3997499 progenyfield pea plant in a field pea plant breeding program comprising:obtaining the field pea plant, or a part thereof, of either cultivar3997499, utilizing said plant, or plant part, as a source of breedingmaterial, and selecting a field pea cultivar 3997499 progeny plant withmolecular markers in common with cultivar 3997499 and/or withmorphological and/or physiological characteristics selected from thecharacteristics listed in Table 1. Breeding steps that may be used inthe field pea plant breeding program include pedigree breeding,backcrossing, mutation breeding, and recurrent selection. In conjunctionwith these steps, techniques such as RFLP-enhanced selection, geneticmarker enhanced selection (for example, SSR markers, SNP markers), andthe making of double haploids may be utilized.

Another method involves producing a population of field pea cultivar3997499 progeny field pea plants, comprising crossing cultivar 3997499with another field pea plant, thereby producing a population of fieldpea plants which, on average, derive 50% of their alleles from field peacultivar 3997499. A plant of this population may be selected andrepeatedly selfed or sibbed with a field pea cultivar resulting fromthese successive filial generations. One embodiment is the field peacultivar produced by this method and that has obtained at least 50% ofits alleles from field pea cultivar 3997499.

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. For example, see, Fehr and Walt, Principles of CultivarDevelopment, pp. 261-286 (1987). Thus, embodiments include field peacultivar 3997499 progeny field pea plants comprising a combination of atleast two cultivar 3997499 traits selected from the group consisting ofthose listed in the Tables or field pea cultivar 3997499 combination oftraits listed in the Summary, so that said progeny field pea plant isnot significantly different for said traits than field pea cultivar3997499 as determined at the 5% significance level when grown in thesame environmental conditions. Using techniques described herein,molecular markers may be used to identify said progeny plant as a fieldpea cultivar 3997499 progeny plant. Mean trait values may be used todetermine whether trait differences are significant, and preferably thetraits are measured on plants grown under the same environmentalconditions. Once such a variety is developed, its value is substantialsince it is important to advance the germplasm base as a whole in orderto maintain or improve traits such as yield, disease resistance, pestresistance, and plant performance in extreme environmental conditions.

Progeny of field pea cultivar 3997499 may also be characterized throughtheir filial relationship with field pea cultivar 3997499, as forexample, being within a certain number of breeding crosses of field peacultivar 3997499. A breeding cross is a cross made to introduce newgenetics into the progeny, and is distinguished from a cross, such as aself or a sib cross, made to select among existing genetic alleles. Thelower the number of breeding crosses in the pedigree, the closer therelationship between field pea cultivar 3997499 and its progeny. Forexample, progeny produced by the methods described herein may be within1, 2, 3, 4, or 5 breeding crosses of field pea cultivar 3997499.

Pedigree Breeding

Pedigree breeding starts with the crossing of two genotypes, such asfield pea cultivar 3997499 and another field pea variety having one ormore desirable characteristics that is lacking or which complementsfield pea cultivar 3997499. If the two original parents do not provideall the desired characteristics, other sources can be included in thebreeding population. In the pedigree method, superior plants are selfedand selected in successive filial generations. In the succeeding filialgenerations, the heterozygous condition gives way to homogeneousvarieties as a result of self-pollination and selection. Typically, inthe pedigree method of breeding, five or more successive filialgenerations of selfing and selection is practiced: F₁ to F₂; F₂ to F₃;F₃ to F₄; F₄ to F₅; etc. After a sufficient amount of inbreeding,successive filial generations will serve to increase seed of thedeveloped variety. Preferably, the developed variety compriseshomozygous alleles at about 95% or more of its loci.

Backcross Breeding

Backcross breeding has been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous cultivaror inbred line which is the recurrent parent. The source of the trait tobe transferred is called the donor parent. After the initial cross,individuals possessing the phenotype of the donor parent are selectedand repeatedly crossed (backcrossed) to the recurrent parent. Theresulting plant is expected to have the attributes of the recurrentparent (e.g., cultivar) and the desirable trait transferred from thedonor parent.

In addition to being used to create a backcross conversion, backcrossingcan also be used in combination with pedigree breeding. As discussedpreviously, backcrossing can be used to transfer one or morespecifically desirable traits from one variety, the donor parent, to adeveloped variety called the recurrent parent, which has overall goodagronomic characteristics yet lacks that desirable trait or traits.However, the same procedure can be used to move the progeny toward thegenotype of the recurrent parent, but at the same time retain manycomponents of the nonrecurrent parent by stopping the backcrossing at anearly stage and proceeding with selfing and selection. For example, afield pea variety may be crossed with another variety to produce afirst-generation progeny plant. The first-generation progeny plant maythen be backcrossed to one of its parent varieties to create a BC₁ orBC₂. Progeny are selfed and selected so that the newly developed varietyhas many of the attributes of the recurrent parent and yet several ofthe desired attributes of the nonrecurrent parent. This approachleverages the value and strengths of the recurrent parent for use in newfield pea varieties.

Therefore, an embodiment is a method of making a backcross conversion offield pea variety 3997499, comprising the steps of crossing a plant offield pea variety 3997499 with a donor plant comprising a desired trait,selecting an Fi progeny plant comprising the desired trait, andbackcrossing the selected Fi progeny plant to a plant of field peavariety 3997499. This method may further comprise the step of obtaininga molecular marker profile of field pea variety 3997499 and using themolecular marker profile to select for a progeny plant with the desiredtrait and the molecular marker profile of field pea cultivar 3997499. Inone embodiment, the desired trait is a mutant gene, gene, or transgenepresent in the donor parent.

Recurrent Selection and Mass Selection

Recurrent selection is a method used in a plant breeding program toimprove a population of plants. Field pea cultivar 3997499 are suitablefor use in a recurrent selection program. The method entails individualplants cross pollinating with each other to form progeny. The progenyare grown and the superior progeny selected by any number of selectionmethods, which include individual plant, half-sib progeny, full-sibprogeny, and selfed progeny. The selected progeny are cross pollinatedwith each other to form progeny for another population. This populationis planted and again superior plants are selected to cross pollinatewith each other. Recurrent selection is a cyclical process and thereforecan be repeated as many times as desired. The objective of recurrentselection is to improve the traits of a population. The improvedpopulation can then be used as a source of breeding material to obtainnew varieties for commercial or breeding use, including the productionof a synthetic cultivar. A synthetic cultivar is the resultant progenyformed by the intercrossing of several selected varieties.

Mass selection is a useful technique when used in conjunction withmolecular marker enhanced selection. In mass selection, seeds fromindividuals are selected based on phenotype or genotype. These selectedseeds are then bulked and used to grow the next generation. Bulkselection requires growing a population of plants in a bulk plot,allowing the plants to self-pollinate, harvesting the seed in bulk, andthen using a sample of the seed harvested in bulk to plant the nextgeneration. Also, instead of self-pollination, directed pollinationcould be used as part of the breeding program.

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified, or created,by intercrossing several different parents. The plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.Single-Seed Descent

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F₂ to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F₂ individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F₂ plantsoriginally sampled in the population will be represented by a progenywhen generation advance is completed.

Multiple-Seed Procedure

In a multiple-seed procedure, breeders commonly harvest one or more podsfrom each plant in a population and thresh them together to form a bulk.Part of the bulk is used to plant the next generation and part is put inreserve. The procedure has been referred to as modified single-seeddescent or the pod-bulk technique.

The multiple-seed procedure has been used to save labor at harvest. Itis considerably faster to thresh pods with a machine than to remove oneseed from each by hand for the single-seed procedure. The multiple-seedprocedure also makes it possible to plant the same number of seeds of apopulation in each generation of inbreeding. Enough seeds are harvestedto make up for those plants that did not germinate or produce seed.

Mutation Breeding

Mutation breeding is another method of introducing new traits into fieldpea variety 3997499. As used herein, a “mutation” is any change in anucleic acid sequence. Nonlimiting examples comprise insertions,deletions, duplications, substitutions, inversions, and translocationsof any nucleic acid sequence, regardless of how the mutation is broughtabout and regardless of how or whether the mutation alters the functionsor interactions of the nucleic acid. For example, and withoutlimitation, a mutation may produce altered enzymatic activity of aribozyme, altered base pairing between nucleic acids (e.g., RNAinterference interactions, DNA-RNA binding, etc.), altered mRNA foldingstability, and/or how a nucleic acid interacts with polypeptides e.g.,DNA-transcription factor interactions, RNA-ribosome interactions,gRNA-endonuclease reactions, etc.). A mutation might result in theproduction of proteins with altered amino acid sequences (e.g., missensemutations, nonsense mutations, frameshift mutations, etc.) and/or theproduction of proteins with the same amino acid sequence ((e.g., silentmutations). Certain synonymous mutations may create no observed changein the plant while others that encode for an identical protein sequencenevertheless result in an altered plant phenotype ((e.g., due to codonusage bias, altered secondary protein structures, etc.). Mutations mayoccur within coding regions (e.g., open reading frames) or outside ofcoding regions (e.g., within promoters, terminators, untranslatedelements, or enhancers), and may affect, for example and withoutlimitation, gene expression levels, gene expression profiles, proteinsequences, and/or sequences encoding RNA elements such as tRNAs,ribozymes, ribosome components, and microRNAs.

Methods disclosed herein are not limited to mutations made in thegenomic DNA of the plant nucleus. For example, in certain embodiments amutation is created in the genomic DNA of an organelle (e.g., a plastidand/or a mitochondrion). In certain embodiments, a mutation is createdin extrachromosomal nucleic acids (including RNA) of the plant, cell, ororganelle of a plant. Nonlimiting examples include creating mutations insupernumerary chromosomes (e.g., B chromosomes), plasmids, and/or vectorconstructs used to deliver nucleic acids to a plant. It is anticipatedthat new nucleic acid forms will be developed and yet fall within thescope of the claimed invention when used with the teachings describedherein.

Methods disclosed herein are not limited to certain techniques ofmutagenesis. Any method of creating a change in a nucleic acid of aplant can be used in conjunction with the disclosed invention, includingthe use of chemical mutagens (e.g. methanesulfonate, sodium azide,aminopurine, etc.), genome/gene editing techniques (e.g. CRISPR-liketechnologies, TALENs, zinc finger nucleases, and meganucleases),ionizing radiation (e.g. ultraviolet and/or gamma rays) temperaturealterations, long-term seed storage, tissue culture conditions,targeting induced local lesions in a genome, sequence-targeted and/orrandom recombinases, etc. It is anticipated that new methods of creatinga mutation in a nucleic acid of a plant will be developed and yet fallwithin the scope of the claimed invention when used with the teachingsdescribed herein.

Similarly, the embodiments disclosed herein are not limited to certainmethods of introducing nucleic acids into a plant and are not limited tocertain forms or structures that the introduced nucleic acids take. Anymethod of transforming a cell of a plant described herein with nucleicacids are also incorporated into the teachings of this innovation, andone of ordinary skill in the art will realize that the use of particlebombardment (e.g. using a gene-gun), Agrobacterium infection and/orinfection by other bacterial species capable of transferring DNA intoplants (e.g., Ochrobactrum sp., Ensifer sp., Rhizobium sp.), viralinfection, and other techniques can be used to deliver nucleic acidsequences into a plant described herein. Methods disclosed herein arenot limited to any size of nucleic acid sequences that are introduced,and thus one could introduce a nucleic acid comprising a singlenucleotide (e.g., an insertion) into a nucleic acid of the plant andstill be within the teachings described herein. Nucleic acids introducedin substantially any useful form, for example, on supernumerarychromosomes (e.g., B chromosomes), plasmids, vector constructs,additional genomic chromosomes (e.g., substitution lines), and otherforms is also anticipated. It is envisioned that new methods ofintroducing nucleic acids into plants and new forms or structures ofnucleic acids will be discovered and yet fall within the scope of theclaimed invention when used with the teachings described herein.

Mutations that occur spontaneously or are artificially induced can beuseful sources of variability for a plant breeder. The goal ofartificial mutagenesis is to increase the rate of mutation for a desiredcharacteristic. Mutation rates can be increased by many different meansincluding temperature, long-term seed storage, tissue cultureconditions, radiation; such as X-rays, Gamma rays (e.g., cobalt 60 orcesium 137), neutrons, (product of nuclear fission by uranium 235 in anatomic reactor), Beta radiation (emitted from radioisotopes such asphosphorus 32 or carbon 14), or ultraviolet radiation (preferably from2500 to 2900 nm), or chemical mutagens (such as base analogues(5-bromo-uracil)), related compounds (8-ethoxy caffeine), antibiotics(streptonigrin), alkylating agents (sulfur mustards, nitrogen mustards,epoxides, ethylenamines, sulfates, sulfonates, sulfones, lactones),azide, hydroxylamine, nitrous acid, or acridines. Once a desired traitis observed through mutagenesis the trait may then be incorporated intoexisting germplasm by traditional breeding techniques. Details ofmutation breeding can be found in Fehr, “Principles of CultivarDevelopment,” Macmillan Publishing Company (1993). In addition,mutations created in other field pea plants may be used to produce abackcross conversion of field pea cultivar 3997499 that comprises suchmutation.

Additional methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct gene transfer method, suchas microprojectile-mediated delivery, DNA injection, electroporation,and the like. More preferably, expression vectors are introduced intoplant tissues by using either microprojectile-mediated delivery with abiolistic device or by using Agrobacterium-mediated transformation.Transformant plants obtained with the protoplasm of the embodiments areintended to be within the scope of the embodiments.

Speed Breeding

Speed breeding uses enhanced lighting (LED) and day-long regimes tooptimize photosynthesis and promote rapid growth of crops. It speeds upthe breeding cycle of plants. For example, six generations of a plantcan be grown per year, compared to two generations using traditionalbreeding methods. Speed breeding can be carried out in numerous ways,one of which involves extending the duration of plants’ daily exposureto light, of up to 22 hours, combined with early seed harvest, to cyclequickly from seed to seed, thereby reducing the generation times forsome long-day or day-neutral crops. Thus, another embodiment includesproducing new field pea cultivars using speed breeding. See for example,Cazzola, Federico, et al., “Speed breeding in pea (Pisum sativum L.), anefficient and simple system to accelerate breeding programs,” Euphytica.October 2020. 216(11): 178 and Gosh, Sreya, et al., “Speed breeding ingrowth chambers and glasshouses for crop breeding and model plantresearch,” Nature Protocols. November 2018. 13(12): 1-20. Thus, anotherembodiment includes using field pea cultivar 3997499 for use in a speedbreeding program.

Gene Editing Using CRISPR

Targeted gene editing can be done using CRISPR/Cas9 technology (Saunders& Joung, Nature Biotechnology, 32, 347-355, 2014). CRISPR is a type ofgenome editing system that stands for Clustered Regularly InterspacedShort Palindromic Repeats. This system and CRISPR-associated (Cas) genesenable organisms, such as select bacteria and archaea, to respond to andeliminate invading genetic material. Ishino, Y., et al. J. Bacteriol.169, 5429-5433 (1987). These repeats were known as early as the 1980 sin E. coli, but Barrangou and colleagues demonstrated that S.thermophilus can acquire resistance against a bacteriophage byintegrating a fragment of a genome of an infectious virus into itsCRISPR locus. Barrangou, R., et al. Science 315, 1709-1712 (2007). Manyplants have already been modified using the CRISPR system, includingfield pea. See for example, Bhowmik, Pankaj, et al., “CRISPR/Cas9 geneediting in legume crops: Opportunities and challenges,” Legume Science.(May 2021). 3(3).

Gene editing can also be done using crRNA-guided surveillance systemsfor gene editing. Additional information about crRNA-guided surveillancecomplex systems for gene editing can be found in the followingdocuments: U.S. Application Publication No. 2010/0076057 (Sontheimer etal., Target DNA Interference with crRNA); U.S. Application PublicationNo. 2014/0179006 (Feng, CRISPR-CAS Component Systems, Methods, andCompositions for Sequence Manipulation); U.S. Application PublicationNo. 2014/0294773 (Brouns et al., Modified Cascade Ribonucleoproteins andUses Thereof); Sorek et al., Annu. Rev. Biochem. 82:273-266, 2013; andWang, S. et al., Plant Cell Rep (2015) 34: 1473-1476.

Therefore, it is another embodiment to use the CRISPR system on fieldpea cultivar 3997499 to modify traits and resistances or tolerances topests, herbicides, and viruses.

Gene Editing Using TALENs

Transcription activator-like effector nucleases (TALENs) have beensuccessfully used to introduce targeted mutations via repair of doublestranded breaks (DSBs) either through nonhomologous end joining (NHEJ),or by homology-directed repair (HDR) and homology-independent repair inthe presence of a donor template. Thus, TALENs are another mechanism fortargeted genome editing using field pea cultivar 3997499. The techniqueis well known in the art; see for example Malzahn, Aimee et al. “Plantgenome editing with TALEN and CRISPR” Cell & bioscience vol. 7 21. 24Apr. 2017.

Therefore, it is another embodiment to use the TALENs system on fieldpea cultivar to modify traits and resistances or tolerances to pests,herbicides, and viruses.

Other Methods of Genome Editing

In addition to CRISPR and TALENs, two other types of engineerednucleases can be used for genome editing: engineered homingendonucleases/meganucleases (EMNs), and zinc finger nucleases (ZFNs).These methods are well known in the art. See for example, Petilino,Joseph F. “Genome editing in plants via designed zinc finger nucleases”In Vitro Cell Dev Biol Plant. 51(1): pp. 1-8 (2015); and Daboussi,Fayza, et al. “Engineering Meganuclease for Precise Plant GenomeModification” in Advances in New Technology for Targeted Modification ofPlant Genomes. Springer Science+Business. pp 21-38 (2015).

Therefore, it is another embodiment to use engineered nucleases on fieldpea cultivar to modify traits and resistances or tolerances to pests,herbicides, and viruses.

Single-Gene Conversions

When the term “field pea plant” or “pea plant” is used in the context ofan embodiment, this also includes any single gene conversions of thatvariety. The term single gene converted plant as used herein refers tothose field pea plants which are developed by a plant breeding techniquecalled backcrossing wherein essentially all of the desired morphologicaland physiological characteristics of a variety are recovered in additionto the single gene transferred into the variety via the backcrossingtechnique. Backcrossing methods can be used with one embodiment toimprove or introduce a characteristic into the variety. The term“backcrossing” as used herein refers to the repeated crossing of ahybrid progeny back to the recurrent parent, i.e., backcrossing 1, 2, 3,4, 5, 6, 7, 8, or more times to the recurrent parent. The parental fieldpea plant that contributes the gene for the desired characteristic istermed the nonrecurrent or donor parent. This terminology refers to thefact that the nonrecurrent parent is used one time in the backcrossprotocol and therefore does not recur. The parental field pea plant towhich the gene or genes from the nonrecurrent parent are transferred isknown as the recurrent parent as it is used for several rounds in thebackcrossing protocol (Poehlman & Sleper (1994); Fehr, Principles ofCultivar Development, pp. 261-286 (1987)). In a typical backcrossprotocol, the original variety of interest (recurrent parent) is crossedto a second variety (nonrecurrent parent) that carries the single geneof interest to be transferred. The resulting progeny from this cross arethen crossed again to the recurrent parent and the process is repeateduntil a field pea plant is obtained wherein essentially all of thedesired morphological and physiological characteristics of the recurrentparent are recovered in the converted plant, in addition to the singletransferred gene from the nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single gene of the recurrent variety ismodified or substituted with the desired gene from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphologicalconstitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross; one ofthe major purposes is to add some agronomically important trait to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered to determine an appropriate testing protocol.Although backcrossing methods are simplified when the characteristicbeing transferred is a dominant allele, a recessive allele may also betransferred. In this instance, it may be necessary to introduce a testof the progeny to determine if the desired characteristic has beensuccessfully transferred.

Many single gene traits have been identified that are not regularlyselected for in the development of a new variety but that can beimproved by backcrossing techniques. Single gene traits may or may notbe transgenic. Examples of these traits include, but are not limited to,male sterility, waxy starch, herbicide resistance, resistance forbacterial, fungal, or viral disease, insect resistance, male fertility,enhanced nutritional quality, industrial usage, yield stability, andyield enhancement. These genes are generally inherited through thenucleus. Several of these single gene traits are described in U.S. Pat.Nos. 5,959,185, 5,973,234, and 5,977,445, the disclosures of which arespecifically hereby incorporated by reference.

Introduction of a New Trait or Locus Into Field Pea Cultivar 3997499

Variety 3997499 represent new varieties into which a new locus or traitmay be introgressed. Direct transformation and backcrossing representtwo important methods that can be used to accomplish such anintrogression. The term backcross conversion and single locus conversionare used interchangeably to designate the product of a backcrossingprogram. Backcross Conversions of Field Pea Cultivar 3997499

A backcross conversion of field pea cultivar 3997499 occurs when DNAsequences are introduced through backcrossing (Hallauer, et al., “CornBreeding,” Corn and Corn Improvements, No. 18, pp. 463-481 (1988)), withfield pea cultivar 3997499 utilized as the recurrent parent. Bothnaturally occurring and transgenic DNA sequences may be introducedthrough backcrossing techniques. A backcross conversion may produce aplant with a trait or locus conversion in at least two or morebackcrosses, including at least 2 crosses, at least 3 crosses, at least4 crosses, at least 5 crosses, and the like. Molecular marker assistedbreeding or selection may be utilized to reduce the number ofbackcrosses necessary to achieve the backcross conversion. For example,see, Openshaw, S. J., et al., Marker-assisted Selection in BackcrossBreeding, Proceedings Symposium of the Analysis of Molecular Data, CropScience Society of America, Corvallis, Oreg. (August 1994), where it isdemonstrated that a backcross conversion can be made in as few as twobackcrosses. In one embodiment, a breeder can combine the teachingsherein with high-density molecular marker profiles spanningsubstantially the entire field pea genome to estimate the value ofselecting certain candidates in a breeding program in a process commonlyknown as genome/genomic selection.

The complexity of the backcross conversion method depends on the type oftrait being transferred (single genes or closely linked genes ascompared to unlinked genes), the level of expression of the trait, thetype of inheritance (cytoplasmic or nuclear), and the types of parentsincluded in the cross. It is understood by those of ordinary skill inthe art that for single gene traits that are relatively easy toclassify, the backcross method is effective and relatively easy tomanage. (See, Hallauer, et al., Corn and Corn Improvement, Sprague andDudley, Third Ed. (1998)). Desired traits that may be transferredthrough backcross conversion include, but are not limited to, sterility(nuclear and cytoplasmic), fertility restoration, nutritionalenhancements, drought tolerance, nitrogen utilization, altered fattyacid profile, low phytate, industrial enhancements, disease resistance(bacterial, fungal, or viral), insect resistance, and herbicideresistance. In addition, an introgression site itself, such as an FRTsite, Lox site, or other site-specific integration site, may be insertedby backcrossing and utilized for direct insertion of one or more genesof interest into a specific plant variety. In some embodiments, thenumber of loci that may be backcrossed into field pea cultivar 3997499is at least 1, 2, 3, 4, or 5, and/or no more than 6, 5, 4, 3, or 2. Asingle locus may contain several transgenes, such as a transgene fordisease resistance that, in the same expression vector, also contains atransgene for herbicide resistance. The gene for herbicide resistancemay be used as a selectable marker and/or as a phenotypic trait. Asingle locus conversion of site-specific integration system allows forthe integration of multiple genes at the converted loci.

The backcross conversion may result from either the transfer of adominant allele or a recessive allele. Selection of progeny containingthe trait of interest is accomplished by direct selection for a traitassociated with a dominant allele. Transgenes transferred viabackcrossing typically function as a dominant single gene trait and arerelatively easy to classify. Selection of progeny for a trait that istransferred via a recessive allele requires growing and selfing thefirst backcross generation to determine which plants carry the recessivealleles. Recessive traits may require additional progeny testing insuccessive backcross generations to determine the presence of the locusof interest. The last backcross generation is usually selfed to givepure breeding progeny for the gene(s) being transferred, although abackcross conversion with a stably introgressed trait may also bemaintained by further backcrossing to the recurrent parent withselection for the converted trait.

Along with selection for the trait of interest, progeny are selected forthe phenotype of the recurrent parent. The backcross is a form ofinbreeding, and the features of the recurrent parent are automaticallyrecovered after successive backcrosses. Poehlman, Breeding Field Crops,p. 204 (1987). Poehlman suggests from one to four or more backcrosses,but as noted above, the number of backcrosses necessary can be reducedwith the use of molecular markers. Other factors, such as a geneticallysimilar donor parent, may also reduce the number of backcrossesnecessary. As noted by Poehlman, backcrossing is easiest for simplyinherited, dominant, and easily recognized traits.

One process for adding or modifying a trait or locus in field peavariety 3997499 comprises crossing field pea cultivar 3997499 plantsgrown from field pea cultivar 3997499 seed with plants of another fieldpea variety that comprise the desired trait or locus, selecting Fiprogeny plants that comprise the desired trait or locus to produceselected Fi progeny plants, crossing the selected progeny plants withthe field pea cultivar 3997499 plants to produce backcross progenyplants, selecting for backcross progeny plants that have the desiredtrait or locus and the morphological characteristics of field peavariety 3997499 to produce selected backcross progeny plants, andbackcrossing to field pea cultivar 3997499 three or more times insuccession to produce selected fourth or higher backcross progeny plantsthat comprise said trait or locus. The modified field pea cultivar3997499 may be further characterized as having the physiological andmorphological characteristics of field pea variety 3997499 listed in theTables as determined at the 5% significance level when grown in the sameenvironmental conditions and/or may be characterized by percentsimilarity or identity to field pea cultivar 3997499 as determined bySSR markers. The above method may be utilized with fewer backcrosses inappropriate situations, such as when the donor parent is highly relatedor markers are used in the selection step. Desired traits that may beused include those nucleic acids known in the art, some of which arelisted herein, that will affect traits through nucleic acid expressionor inhibition. Desired loci include the introgression of FRT, Lox, andother sites for site specific integration, which may also affect adesired trait if a functional nucleic acid is inserted at theintegration site.

In addition, the above process and other similar processes describedherein may be used to produce first generation progeny field pea seed byadding a step at the end of the process that comprises crossing fieldpea cultivar 3997499 with the introgressed trait or locus with adifferent field pea plant and harvesting the resultant first-generationprogeny field pea seed.

Molecular Techniques Using Field Pea Cultivar 3997499

The advent of new molecular biological techniques has allowed theisolation and characterization of genetic elements with specificfunctions, such as encoding specific protein products. Scientists in thefield of plant biology developed a strong interest in engineering thegenome of plants to contain and express foreign genetic elements, oradditional, or modified versions of native or endogenous geneticelements in order to “alter” (the utilization of up-regulation,down-regulation, or gene silencing) the traits of a plant in a specificmanner. Any DNA sequences, whether from a different species or from thesame species, which are introduced into the genome using transformationor various breeding methods are referred to herein collectively as“transgenes.” In some embodiments, a transgenic variant of field peacultivar 3997499 may contain at least one transgene but could contain atleast 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and/or no more than 15, 14, 13, 12,11, 10, 9, 8, 7, 6, 5, 4, 3, or 2. Over the last fifteen to twenty yearsseveral methods for producing transgenic plants have been developed, andanother embodiment also relates to transgenic variants of the claimedfield pea variety 3997499.

Nucleic acids or polynucleotides refer to RNA or DNA that is linear orbranched, single or double stranded, or a hybrid thereof. The term alsoencompasses RNA/DNA hybrids. These terms also encompass untranslatedsequence located at both the 3′ and 5′ ends of the coding region of thegene: at least about 1000 nucleotides of sequence upstream from the 5′end of the coding region and at least about 200 nucleotides of sequencedownstream from the 3′ end of the coding region of the gene. Less commonbases, such as inosine, 5-methylcytosine, 6-methyladenine, hypoxanthineand others can also be used for antisense, dsRNA and ribozyme pairing.For example, polynucleotides that contain C-5 propyne analogues ofuridine and cytidine have been shown to bind RNA with high affinity andto be potent antisense inhibitors of gene expression. Othermodifications, such as modification to the phosphodiester backbone, orthe 2′-hydroxy in the ribose sugar group of the RNA can also be made.The antisense polynucleotides and ribozymes can consist entirely ofribonucleotides, or can contain mixed ribonucleotides anddeoxyribonucleotides. The polynucleotides of the embodiments may beproduced by any means, including genomic preparations, cDNApreparations, in-vitro synthesis, RT-PCR, and in vitro or in vivotranscription.

One embodiment is a process for producing field pea variety 3997499further comprising a desired trait, said process comprising introducinga transgene that confers a desired trait to a field pea plant of variety3997499. Another embodiment is the product produced by this process. Inone embodiment, the desired trait may be one or more of herbicideresistance, insect resistance, disease resistance, decreased phytate, ormodified fatty acid or carbohydrate metabolism. The specific gene may beany known in the art or listed herein, including: a polynucleotideconferring resistance to imidazolinone, dicamba, sulfonylurea,glyphosate, glufosinate, triazine, PPO-inhibitor herbicides,benzonitrile, cyclohexanedione, phenoxy proprionic acid, andL-phosphinothricin; a polynucleotide encoding a Bacillus thuringiensispolypeptide; a polynucleotide encoding phytase, FAD-2, FAD-3, galactinolsynthase, or a raffinose synthetic enzyme; or a polynucleotideconferring resistance to root rot, stem rot, or Phytophthora root rot.

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. See, forexample, Miki et al., “Procedures for Introducing Foreign DNA intoPlants,” in Methods in Plant Molecular Biology and Biotechnology, Glickand Thompson Eds., CRC Press, Inc., Boca Raton, pp. 67-88 (1993), andArmstrong, “The First Decade of Maize Transformation: A Review andFuture Perspective,” Maydica, 44:101-109 (1999). In addition, expressionvectors and in vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available. See, forexample, Esposito, M.A. et al., “A rapid method to increase the numberof F₁ plants in pea (Pisum sativum) breeding programs,” Genet Mol Res.2012 Aug 16;11(3): 2729-32; Gruber, et al., “Vectors for PlantTransformation,” in Methods in Plant Molecular Biology andBiotechnology, Glick and Thompson Eds., CRC Press, Inc., Boca Raton, pp.89-119 (1993).

A genetic trait which has been engineered into the genome of aparticular field pea plant may then be moved into the genome of anothervariety using traditional breeding techniques that are well-known in theplant breeding arts. For example, a backcrossing approach is commonlyused to move a transgene from a transformed field pea variety into analready developed field pea variety, and the resulting backcrossconversion plant would then comprise the transgene(s).

Various genetic elements can be introduced into the plant genome usingtransformation. These elements include, but are not limited to, genes,coding sequences, inducible, constitutive and tissue specific promoters,enhancing sequences, and signal and targeting sequences. For example,see the traits, genes, and transformation methods listed in U.S. Pat.No. 6,118,055. Breeding with Molecular Markers

Molecular markers, which includes markers identified through the use oftechniques such as Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats(SSRs), and Single Nucleotide Polymorphisms (SNPs), may be used in plantbreeding methods utilizing field pea cultivar 3997499.

Isozyme Electrophoresis and RFLPs have been widely used to determinegenetic composition. Jacobsen, Hans-Jörg, “Analysis of RNase isozymes ingerminating pea cotyledons by polyacrylamide-gel electrophoresis,” Plantand Cell Physiology, June 1980, 21(4):659-665; Dirlewanger, E. et al.,“Restriction fragment length polymorphism analysis of loci associatedwith disease resistance genes and developmental traits in Pisum sativumL,” Theor Appl Genet. 1994. 88(1):17-27; Dhillon, N.P.S., et al.,“Isozyme and RFLP mapping of sbm-4, a gene in pea (Pisum sativum)conferring resistance to the P-4 pathotype of pea seed borne mosaicvirus,” Adv. Hort. Sci. (1995). 9:159-161.

SSR technology is also an efficient and practical marker technology.More marker loci can be routinely used, and more alleles per markerlocus can be found, using SSRs in comparison to RFLPs. See for example,Xuelian, Sun, et al., “SSR genetic linkage map construction of pea(Pisum sativum L .) based on Chinese native varieties,” The CropJournal. (April-June 2014). 2(2-3): 170-174; Tao, Yang, et al.,“High-Throughput Development of SSR Markers from Pea (Pisum sativum L .)Based on Next Generation Sequencing of a Purified Chinese CommercialVariety,” PLoS One. (2015). 10(10): e0139775; Hanci, Faith, “Geneticvariability in peas (Pisum sativum L.) from Turkey assessed withmolecular and morphological markers,” Folia Hort. (2019). 31(1):101-116. Single Nucleotide Polymorphisms may also be used to identifythe unique genetic composition of the embodiment(s) and progenyvarieties retaining that unique genetic composition. Various molecularmarker techniques may be used in combination to enhance overallresolution.

Field pea DNA molecular marker linkage maps have been rapidlyconstructed and widely implemented in genetic studies. See for example,Ellis, T., et al., “Linkage Maps in Pea,” Genetics. (1992 Mar). 130(3):649-663; Xuelian, Sun, et al., “SSR genetic linkage map construction ofpea (Pisum sativum L .) based on Chinese native varieties,” The CropJournal. (April-June 2014). 2(2-3): 170-174; and Gilpin, B.J., et al.,“A linkage map of the pea (Pisum sativum L.) genome containing clonedsequences of known function and expressed sequence tags (ESTs),”Theoretical and Applied Genetics. (1997). 95: 1289-1299.

Quantitative trait loci (QTL) refer to genetic loci that control to somedegree numerically representable traits that are usually continuouslydistributed. One use of molecular markers is Quantitative Trait Loci(QTL) mapping. QTL mapping is the use of markers, which are known to beclosely linked to alleles that have measurable effects on a quantitativetrait. Selection in the breeding process is based upon the accumulationof markers linked to the positive effecting alleles and/or theelimination of the markers linked to the negative effecting alleles fromthe plant’s genome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select for the genome of the recurrent parent and against thegenome of the donor parent. Using this procedure can minimize the amountof genome from the donor parent that remains in the selected plants. Itcan also be used to reduce the number of crosses back to the recurrentparent needed in a backcrossing program. The use of molecular markers inthe selection process is often called genetic marker enhanced selection.Molecular markers may also be used to identify and exclude certainsources of germplasm as parental varieties or ancestors of a plant byproviding a means of tracking genetic profiles through crosses.

Production of Double Haploids

The production of double haploids can also be used for the developmentof plants with a homozygous phenotype in the breeding program. Forexample, a field pea plant for which field pea cultivar 3997499 is aparent can be used to produce double haploid plants. Double haploids areproduced by the doubling of a set of chromosomes (1N) from aheterozygous plant to produce a completely homozygous individual. Forexample, see, Deswal, Kapil, “Progress and opportunities in doublehaploid production in lentil (Lens culinaris Medik.), soybean (Glycinemax L. Merr.) and chickpea (Cicer arietinum L .),” Journal ofPharmacognosy and Phytochemistry. (2018). 7(3): 3105-3109 and U.S. Pat.No. 7,135,615. This can be advantageous because the process omits thegenerations of selfing needed to obtain a homozygous plant from aheterozygous source.

Haploid induction systems have been developed for various plants toproduce haploid tissues, plants and seeds. The haploid induction systemcan produce haploid plants from any genotype by crossing a selected line(as female) with an inducer line. Such inducer lines for maize includeStock 6 (Coe, Am. Nat., 93:381-382 (1959); Sharkar and Coe, Genetics,54:453-464 (1966); KEMS (Deimling, Roeber, and Geiger, Vortr.Pflanzenzuchtg, 38:203-224 (1997); or KMS and ZMS (Chalyk, Bylich &Chebotar, MNL, 68:47 (1994); Chalyk & Chebotar, Plant Breeding,119:363-364 (2000)); and indeterminate gametophyte (ig) mutation(Kermicle, Science, 166:1422-1424 (1969)). The disclosures of which areincorporated herein by reference.

Methods for obtaining haploid plants are also disclosed in Kobayashi,M., et al., Journ. of Heredity, 71(1):9-14 (1980); Pollacsek, M.,Agronomie (Paris) 12(3):247-251 (1992); Cho-Un-Haing, et al., Journ. ofPlant Biol., 39(3): 185-188 (1996); Verdoodt, L., et al., 96(2):294-300(February 1998); Chalyk, et al., Maize Genet Coop., Newsletter 68:47(1994).

Thus, an embodiment is a process for making a substantially homozygousfield pea cultivar 3997499 progeny plant by producing or obtaining aseed from the cross of field pea cultivar 3997499 and another field peaplant and applying double haploid methods to the Fi seed or Fi plant orto any successive filial generation.

A process of making seed retaining the molecular marker profile of fieldpea variety 3997499 is contemplated, such process comprising obtainingor producing Fi seed for which field pea variety 3997499 is a parent,inducing doubled haploids to create progeny without the occurrence ofmeiotic segregation, obtaining the molecular marker profile of fieldvariety 3997499, and selecting progeny that retain the molecular markerprofile of field pea cultivar 3997499.

Production of Apomictic Plants

Apomixis is a naturally occurring mode of asexual reproduction inflowering plants. Apomixis results in genetic identity of the offspringof a mother plant. This process results in seed formation without theinvolvement of meiosis or fertilization of the egg. Apomictic processesbypass meiosis and fertilization, leading directly to clonal embryoformation. The mother plant can be highly heterozygous, but sinceapomixis bypasses meiosis, there is no segregation of traits amongseed-derived progeny. Apomictic hybrids are true-breeding hybridsbecause seed-derived progeny of an apomictic plant are geneticallyidentical to the maternal parent. In other words, apomictic hybrids areclonal in origin. Apomixis is characterized by: 1) apomeiosis, whichrefers to the formation of unreduced embryo sacs derived from nucellarcells of the ovary, and 2) parthenogenesis, which refers to thedevelopment of the unreduced egg into an embryo. Many types of plantspecies feature apomictic reproduction and can be propagated asexually.Apomixis can be used for more efficient hybrid seed production in hybridcrops, eliminating the need to use separate male and female parentsgrown in isolation to generate hybrid seed. Apomixis can also be usedfor seed propagation of heterozygous crops that typically arevegetatively propagated through tubers, organs that can harbor andtransmit diseases across generations. Apomixis can also promote thedevelopment of hybrids in crops where hybrids currently are notavailable due to the lack of parental lines that can be easily crossedon a commercial scale. Apomixis can be used as a breeding tool toincrease and test large numbers of novel hybrids generated by sexualreproduction, but increased through apomictic reproduction. Thus, oneembodiment includes egg development into an embryo withoutfertilization, as an essential component of apomixis or clonalreproduction through seeds and further, plants of the presentembodiments can be part of or generated from a breeding program, and mayalso be reproduced using apomixis. Methods for the production ofapomictic plants are known in the art. See for example, U.S. Pat. Nos.10,633,672, 10,954,525, 10,907,174, and 5,811,636. Thus, one embodimentincludes using field pea variety 3997499 as a source material forcreating apomictic plants.

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks. See for example, Biddle, A.J., “Peas and Beans,” (2017). CABI.Biddle, A.J., Editor. Chapter 3, page 38; Allard (1960); Simmonds(1979); Sneep, et al. (1979); Fehr (1987).

Expression Vectors for Field Pea Transformation: Marker Genes

Plant transformation involves the construction of an expression vectorwhich will function in plant cells. Such a vector comprises DNAcomprising a gene under control of, or operatively linked to, aregulatory element (for example, a promoter). Expression vectors includeat least one genetic marker operably linked to a regulatory element (forexample, a promoter) that allows transformed cells containing the markerto be either recovered by negative selection, i.e., inhibiting growth ofcells that do not contain the selectable marker gene, or by positiveselection, i.e., screening for the product encoded by the geneticmarker. Many commonly used selectable marker genes for planttransformation are well-known in the transformation arts, and include,for example, genes that code for enzymes that metabolically detoxify aselective chemical agent which may be an antibiotic or an herbicide, orgenes that encode an altered target which is insensitive to theinhibitor. A few positive selection methods are also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) gene which, when under thecontrol of plant regulatory signals, confers resistance to kanamycin.Fraley, et al., Proc. Natl. Acad. Sci. USA, 80:4803 (1983); Svabova, L.,et al., “Agrobacterium-mediated transformation of Pisum sativum in vitroand in vivo,” Biologia Plantarum. (2005). 49: 361-370.

Another commonly used selectable marker gene is the hygromycinphosphotransferase gene which confers resistance to the antibiotichygromycin. Vanden Elzen, et al., Plant Mol. Biol., 5:299 (1985).

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase and aminoglycoside-3′adenyl transferase,the bleomycin resistance determinant (Hayford, et al., Plant Physiol.,86:1216 (1988); Jones, et al., Mol. Gen. Genet., 210:86 (1987); Svab, etal., Plant Mol. Biol., 14:197 (1990); Hille, et al., Plant Mol. Biol.,7:171 (1986)). Other selectable marker genes confer resistance toherbicides such as glyphosate, glufosinate, or bromoxynil (Comai, etal., Nature, 317:741-744 (1985); Gordon-Kamm, et al., Plant Cell,2:603-618 (1990); Stalker, et al., Science, 242:419-423 (1988)).

Selectable marker genes for plant transformation not of bacterial origininclude, for example, mouse dihydrofolate reductase, plant5-enolpyruvylshikimate-3-phosphate synthase, and plant acetolactatesynthase (Eichholtz, et al., Somatic Cell Mol. Genet., 13:67 (1987);Shah, et al., Science, 233:478 (1986); Charest, et al., Plant Cell Rep.,8:643 (1990)).

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells, rather than directgenetic selection of transformed cells, for resistance to a toxicsubstance such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used genes for screeningpresumptively transformed cells include β-glucuronidase (GUS),β-galactosidase, luciferase, and chloramphenicol acetyltransferase(Jefferson, R. A., Plant Mol. Biol. Rep., 5:387 (1987); Teeri, et al.,EMBO J., 8:343 (1989); Koncz, et al., Proc. Natl. Acad. Sci. USA, 84:131(1987); DeBlock, et al., EMBO J., 3:1681 (1984)).

In vivo methods for visualizing GUS activity that do not requiredestruction of plant tissue are available (Molecular Probes, Publication2908, IMAGENE GREEN, pp. 1-4 (1993); Naleway, et al., J. Cell Biol.,115:151a (1991)). However, these in-vivo methods for visualizing GUSactivity have not proven useful for recovery of transformed cellsbecause of low sensitivity, high fluorescent backgrounds, andlimitations associated with the use of luciferase genes as selectablemarkers.

A gene encoding Green Fluorescent Protein (GFP) has been utilized as amarker for gene expression in prokaryotic and eukaryotic cells. See Liu,A.X., et al., “Soluble expression and characterization of a GFP-fusedpea actin isoform (PEAcl),” Cell Research. (2004). 14: 407-414; andChalfie, et al., Science, 263:802 (1994). GFP and mutants of GFP may beused as screenable markers.

Expression Vectors for Field Pea Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element (for example, a promoter).Several types of promoters are well known in the transformation arts asare other regulatory elements that can be used alone or in combinationwith promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred.”Promoters that initiate transcription only in a certain tissue arereferred to as “tissue-specific.” A “cell-type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may affect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell-type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter that is active under mostenvironmental conditions.

A. Inducible Promoters: An inducible promoter is operably linked to agene for expression in field pea. Optionally, the inducible promoter isoperably linked to a nucleotide sequence encoding a signal sequencewhich is operably linked to a gene for expression in field pea. With aninducible promoter, the rate of transcription increases in response toan inducing agent.

Any inducible promoter can be used in an embodiment(s). See, Ward, etal., Plant Mol. Biol., 22:361-366 (1993). Exemplary inducible promotersinclude, but are not limited to, that from the ACEI system whichresponds to copper (Mett, et al., Proc. Natl. Acad. Sci. USA,90:4567-4571 (1993)); In2 gene from maize which responds tobenzenesulfonamide herbicide safeners (Hershey, et al., Mol. GenGenetics, 227:229-237 (1991); Gatz, et al., Mol. Gen. Genetics,243:32-38 (1994)); or Tet repressor from Tn10 (Gatz, et al., Mol. Gen.Genetics, 227:229-237 (1991)). An inducible promoter is a promoter thatresponds to an inducing agent to which plants do not normally respond.An exemplary inducible promoter is the inducible promoter from a steroidhormone gene, the transcriptional activity of which is induced by aglucocorticosteroid hormone (Schena, et al., Proc. Natl. Acad. Sci. USA,88:0421 (1991)).

B. Constitutive Promoters: A constitutive promoter is operably linked toa gene for expression in field pea or the constitutive promoter isoperably linked to a nucleotide sequence encoding a signal sequencewhich is operably linked to a gene for expression in field pea.

Many different constitutive promoters can be utilized in anembodiment(s). Exemplary constitutive promoters include, but are notlimited to, the promoters from plant viruses such as the 35S promoterfrom CaMV (Odell, et al., Nature, 313:810-812 (1985)) and the promotersfrom such genes as rice actin (McElroy, et al., Plant Cell, 2:163-171(1990)); ubiquitin (Christensen, et al., Plant Mol. Biol., 12:619-632(1989); Christensen, et al., Plant Mol. Biol., 18:675-689 (1992)); pEMU(Last, et al., Theor. Appl. Genet., 81:581-588 (1991)); MAS (Velten, etal., EMBO J., 3:2723-2730 (1984)); and maize H3 histone (Lepetit, etal., Mol. Gen. Genetics, 231:276-285 (1992); Atanassova, et al., PlantJournal, 2 (3):291-300 (1992)). The ALS promoter, Xbal/NcoI fragment 5′to the Brassica napus ALS3 structural gene (or a nucleotide sequencesimilarity to said XbaI/NcoI fragment), represents a particularly usefulconstitutive promoter. See, U.S. Pat. No. 5,659,026.

C. Tissue-Specific or Tissue-Preferred Promoters: A tissue-specificpromoter is operably linked to a gene for expression in field pea.Optionally, the tissue-specific promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in field pea. Plants transformed with a gene ofinterest operably linked to a tissue-specific promoter produce theprotein product of the transgene exclusively, or preferentially, in aspecific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in anembodiment(s). Exemplary tissue-specific or tissue-preferred promotersinclude, but are not limited to, a rootpreferred promoter such as thatfrom the phaseolin gene (Murai, et al., Science, 23:476-482 (1983);Sengupta-Gopalan, et al., Proc. Natl. Acad. Sci. USA, 82:3320-3324(1985)); a leafspecific and light-induced promoter such as that from cabor rubisco (Simpson, et al., EMBO J., 4(11):2723-2729 (1985); Timko, etal., Nature, 318:579-582 (1985)); an anther-specific promoter such asthat from LAT52 (Twell, et al., Mol. Gen. Genetics, 217:240-245 (1989));a pollen-specific promoter such as that from Zm13 (Guerrero, et al.,Mol. Gen. Genetics, 244:161-168 (1993)); or a microspore-preferredpromoter such as that from apg (Twell, et al., Sex. Plant Reprod.,6:217-224 (1993)).

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of a protein produced by transgenes to a subcellularcompartment, such as the chloroplast, vacuole, peroxisome, glyoxysome,cell wall, or mitochondrion, or for secretion into the apoplast, isaccomplished by means of operably linking the nucleotide sequenceencoding a signal sequence to the 5′ and/or 3′ region of a gene encodingthe protein of interest. Targeting sequences at the 5′ and/or 3′ end ofthe structural gene may determine during protein synthesis andprocessing where the encoded protein is ultimately compartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample, Becker, et al., Plant Mol. Biol., 20:49 (1992); Knox, C., etal., Plant Mol. Biol., 9:3-17 (1987); Lerner, et al., Plant Physiol.,91:124-129 (1989); Frontes, et al., Plant Cell, 3:483-496 (1991);Matsuoka, et al., Proc. Natl. Acad. Sci., 88:834 (1991); Gould, et al.,J. Cell. Biol., 108:1657 (1989); Creissen, et al., Plant J., 2:129(1991); Kalderon, et al., Cell, 39:499-509 (1984); Steifel, et al.,Plant Cell, 2:785-793 (1990).

Foreign Protein Genes and Agronomic Genes: Transformation

With transgenic plants according to one embodiment, a foreign proteincan be produced in commercial quantities. Thus, techniques for theselection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein can then beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Emkani, M., et al., “Pea ProteinExtraction Assisted by Lactic Fermentation: Impact on Protein Profileand Thermal Properties,” Foods. (2021). 10(3): 549; and Heney and Orr,Anal. Biochem., 114:92-6 (1981).

According to an embodiment, the transgenic plant provided for commercialproduction of foreign protein is a field pea plant. In anotherembodiment, the biomass of interest is seed. For the relatively smallnumber of transgenic plants that show higher levels of expression, agenetic map can be generated, primarily via conventional RFLP, PCR, andSSR analysis, which identifies the approximate chromosomal location ofthe integrated DNA molecule. For exemplary methodologies in this regard,see, Glick and Thompson, Methods in Plant Molecular Biology andBiotechnology, CRC Press, Inc., Boca Raton, 269:284 (1993). Mapinformation concerning chromosomal location is useful for proprietaryprotection of a subject transgenic plant.

Map information concerning chromosomal location is useful forproprietary protection of a subject transgenic plant. If unauthorizedpropagation is undertaken and crosses made with other germplasm, the mapof the integration region can be compared to similar maps for suspectplants to determine if the latter have a common parentage with thesubject plant. Map comparisons would involve hybridizations, RFLP, PCR,SSR, and sequencing, all of which are conventional techniques. SNPs mayalso be used alone or in combination with other techniques. See forexample, Leonforte, Antonio, et al., “SNP marker discovery, linkage mapconstruction and identification of QTLs for enhanced salinity tolerancein field pea (Pisum sativum L .),” BMC Plant Biol. (2013). 13: 161; andSudheesh, Shimha, et al., “Consensus Genetic Map Construction for FieldPea (Pisum sativum L .), Trait Dissection of Biotic and Abiotic StressTolerance and Development of a Diagnostic Marker for the er1 PowderyMildew Resistance Gene,” (December 2014). Plant Molecular BiologyReporter. 33(5).

Likewise, by means of one embodiment, plants can be geneticallyengineered to express various phenotypes of agronomic interest. Throughthe transformation of field pea, the expression of genes can be alteredto enhance disease resistance, insect resistance, herbicide resistance,agronomic, grain quality, and other traits. Transformation can also beused to insert DNA sequences which control or help controlmale-sterility. DNA sequences native to field pea, as well as non-nativeDNA sequences, can be transformed into field pea and used to alterlevels of native or non-native proteins. Various promoters, targetingsequences, enhancing sequences, and other DNA sequences can be insertedinto the genome for the purpose of altering the expression of proteins.The interruption or suppression of the expression of a gene at the levelof transcription or translation (also known as gene silencing or genesuppression) is desirable for several aspects of genetic engineering inplants.

Many techniques for gene silencing are well-known to one of skill in theart, including, but not limited to, knock-outs (such as by insertion ofa transposable element such as Mu (Vicki Chandler, The Maize Handbook,Ch. 118 (Springer-Verlag 1994)) or other genetic elements such as a FRT,Lox, or other site specific integration sites; antisense technology(see, i.e., Sheehy, et al., PNAS USA, 85:8805-8809 (1988) and U.S. Pat.Nos. 5,107,065, 5,453,566, and 5,759,829); co-suppression (i.e., Taylor,Plant Cell, 9:1245 (1997); Jorgensen, Trends Biotech., 8(12):340-344(1990); Flavell, PNAS USA, 91:3490-3496 (1994); Finnegan, et al.,Bio/Technology, 12:883-888 (1994); Neuhuber, et al., Mol. Gen. Genet.,244:230-241 (1994)); RNA interference (Napoli, et al., Plant Cell,2:279-289 (1990); U.S. Pat. No. 5,034,323; Sharp, Genes Dev., 13:139-141(1999); Zamore, et al., Cell, 101:25-33 (2000); Montgomery, et al., PNASUSA, 95:15502-15507 (1998)), virus-induced gene silencing (Burton, etal., Plant Cell, 12:691-705 (2000); Baulcombe, Curr. Op. Plant Bio.,2:109-113 (1999)); target-RNA-specific ribozymes (Haseloff, et al.,Nature, 334:585-591 (1988)); hairpin structures (Smith, et al., Nature,407:319-320 (2000); U.S. Pat. Nos. 6,423,885, 7,138,565, 6,753,139, and7,713,715); MicroRNA (Aukerman & Sakai, Plant Cell, 15:2730-2741(2003)); ribozymes (Steinecke, et al., EMBO J., 11:1525 (1992);Perriman, et al., Antisense Res. Dev., 3:253 (1993)); oligonucleotidemediated targeted modification (i.e., U.S. Pat. Nos. 6,528,700 and6,911,575); Zn-finger targeted molecules (i.e., U.S. Pat. Nos.7,151,201, 6,453,242, 6,785,613, 7,177,766 and 7,788,044); and othermethods or combinations of the above methods known to those of skill inthe art.

Methods for Field Pea Transformation

Numerous methods for plant transformation have been developed includingbiological and physical plant transformation protocols. See, forexample, Miki, et al., “Procedures for Introducing Foreign DNA intoPlants,” in Methods in Plant Molecular Biology and Biotechnology, Glickand Thompson Eds., CRC Press, Inc., Boca Raton, pp. 67-88 (1993). Inaddition, expression vectors and in-vitro culture methods for plant cellor tissue transformation and regeneration of plants are available. See,for example, Gruber, et al., “Vectors for Plant Transformation,” inMethods in Plant Molecular Biology and Biotechnology, Glick and ThompsonEds., CRC Press, Inc., Boca Raton, pp. 89-119 (1993).

A. Agrobacterium-mediated Transformation: One method for introducing anexpression vector into plants is based on the natural transformationsystem of Agrobacterium. See, for example, Horsch, et al., Science,227:1229 (1985). A. tumefaciens and A. rhizogenes are plant pathogenicsoil bacteria which genetically transform plant cells. The Ti and Riplasmids of A. tumefaciens and A. rhizogenes, respectively, carry genesresponsible for genetic transformation of the plant. See, for example,Kado, C. I., Crit. Rev. Plant Sci., 10:1 (1991). Descriptions ofAgrobacterium vector systems and methods for Agrobacterium-mediated genetransfer are provided by Gruber, et al., supra, Miki, et al., supra, andMoloney, et al., Plant Cell Reports, 8:238 (1989). See also, U.S. Pat.No. 5,563,055 (Townsend and Thomas), issued Oct. 8, 1996.

B. Direct Gene Transfer: Several methods of plant transformation,collectively referred to as direct gene transfer, have been developed asan alternative to Agrobacterium-mediated transformation. A generallyapplicable method of plant transformation is microprojectile-mediatedtransformation where DNA is carried on the surface of microprojectilesmeasuring 1 to 4 µm. The expression vector is introduced into planttissues with a biolistic device that accelerates the microprojectiles tospeeds of 300 to 600 m/s which is sufficient to penetrate plant cellwalls and membranes. Sanford, et al., Part. Sci. Technol., 5:27 (1987);Sanford, J. C., Trends Biotech., 6:299 (1988); Klein, et al., Bio/Tech.,6:559-563 (1988); Sanford, J. C., Physiol Plant, 7:206 (1990); Klein, etal., Biotechnology, 10:268 (1992). See also, U.S. Pat. No. 5,015,580(Christou, et al.), issued May 14, 1991 and U.S. Pat. No. 5,322,783.

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang, et al., Bio/Technology, 9:996 (1991).Alternatively, liposome and spheroplast fusion have been used tointroduce expression vectors into plants. Deshayes, et al., EMBO J.,4:2731 (1985); Christou, et al., Proc Natl. Acad. Sci. USA, 84:3962(1987). Direct uptake of DNA into protoplasts using CaCl₂ precipitation,polyvinyl alcohol or poly-L-ornithine have also been reported. Hain, etal., Mol. Gen. Genet., 199:161 (1985) and Draper, et al., Plant CellPhysiol., 23:451 (1982). Electroporation of protoplasts and whole cellsand tissues has also been described (D'Halluin, et al., Plant Cell,4:1495-1505 (1992); and Spencer, et al., Plant Mol. Biol., 24:51-61(1994)).

Following transformation of field pea target tissues, expression of theabove-described selectable marker genes allows for preferentialselection of transformed cells, tissues, and/or plants, usingregeneration and selection methods well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic variety. The transgenic variety could then becrossed with another (non-transformed or transformed) variety in orderto produce a new transgenic variety. Alternatively, a genetic trait thathas been engineered into a particular field pea line using the foregoingtransformation techniques could be moved into another line usingtraditional backcrossing techniques that are well known in the plantbreeding arts. For example, a backcrossing approach could be used tomove an engineered trait from a public, non-elite variety into an elitevariety, or from a variety containing a foreign gene in its genome intoa variety or varieties that do not contain that gene. As used herein,“crossing” can refer to a simple x by y cross or the process ofbackcrossing depending on the context.

Likewise, by means of one embodiment, agronomic genes can be expressedin transformed plants. More particularly, plants can be geneticallyengineered to express various phenotypes of agronomic interest.Exemplary genes implicated in this regard include, but are not limitedto, those categorized below:

-   1. Genes That Confer Resistance to Pests or Disease and That Encode:    -   A. Plant disease resistance genes. Plant defenses are often        activated by specific interaction between the product of a        disease resistance gene (R) in the plant and the product of a        corresponding avirulence (Avr) gene in the pathogen. A plant        variety can be transformed with one or more cloned resistance        genes to engineer plants that are resistant to specific pathogen        strains. See, for example, Jones, et al., Science,        266:789 (1994) (cloning of the tomato Cf-9 gene for resistance        to Cladosporium fulvum); Martin, et al., Science,        262:1432 (1993) (tomato Pto gene for resistance to Pseudomonas        syringae pv. tomato encodes a protein kinase); Mindrinos, et        al., Cell, 78:1089 (1994) (Arabidopsis RSP2 gene for resistance        to Pseudomonas syringae); McDowell & Woffenden, Trends        Biotechnol., 21(4):178-83 (2003); and Toyoda, et al., Transgenic        Res., 11 (6):567-82 (2002).    -   B. A gene conferring resistance to a pest, such as gram pod        borer. See, Singh, Shweta, et al., “Expression of Cry2Aa, a        Bacillus thuringiensis insecticidal protein in transgenic pigeon        pea confers resistance to gram pod borer, Helicoverpa armiger,”        Scientific Reports. (2018). 8: Article number: 8820.    -   C. A Bacillus thuringiensis protein, a derivative thereof or a        synthetic polypeptide modeled thereon. See, for example, Geiser,        et al., Gene, 48:109 (1986), who disclose the cloning and        nucleotide sequence of a Bt δ-endotoxin gene. Moreover, DNA        molecules encoding δ-endotoxin genes can be purchased from        American Type Culture Collection, Manassas, Va., for example,        under ATCC Accession Nos. 40098, 67136, 31995, and 31998.    -   D. A lectin. See, for example, Van Damme, et al., Plant Molec.        Biol., 24:25 (1994), who disclose the nucleotide sequences of        several Clivia miniata mannose-binding lectin genes.    -   E. A vitamin-binding protein such as avidin. See, International        Application No. PCT/US1993/006487, which teaches the use of        avidin and avidin homologues as larvicides against insect pests.    -   F. An enzyme inhibitor, for example, a protease or proteinase        inhibitor or an amylase inhibitor. See, for example, Abe, et        al., J. Biol. Chem., 262:16793 (1987) (nucleotide sequence of        rice cysteine proteinase inhibitor); Huub, et al., Plant Molec.        Biol., 21:985 (1993) (nucleotide sequence of cDNA encoding        tobacco proteinase inhibitor I); Sumitani, et al., Biosci.        Biotech. Biochem., 57:1243 (1993) (nucleotide sequence of        Streptomyces nitrosporeus α-amylase inhibitor); and U.S. Pat.        No. 5,494,813.    -   G. An insect-specific hormone or pheromone, such as an        ecdysteroid or juvenile hormone, a variant thereof, a mimetic        based thereon, or an antagonist or agonist thereof. See, for        example, the disclosure by Hammock, et al., Nature, 344:458        (1990), of baculovirus expression of cloned juvenile hormone        esterase, an inactivator of juvenile hormone.    -   H. An insect-specific peptide or neuropeptide which, upon        expression, disrupts the physiology of the affected pest. For        example, see the disclosures of Regan, J. Biol. Chem.,        269:9 (1994) (expression cloning yields DNA coding for insect        diuretic hormone receptor); Pratt, et al., Biochem. Biophys.        Res. Comm., 163:1243 (1989) (an allostatin is identified in        Diploptera puntata); Chattopadhyay, et al., Critical Reviews in        Microbiology, 30(1):33-54 (2004); Zjawiony, J. Nat. Prod.,        67(2):300-310 (2004); Carlini & Grossi-de-Sa, Toxicon,        40(11):1515-1539 (2002); Ussuf, et al., Curr Sci., 80(7):847-853        (2001); Vasconcelos & Oliveira, Toxicon, 44(4):385-403 (2004).        See also, U.S. Pat. No. 5,266,317 which discloses genes encoding        insect-specific, paralytic neurotoxins.    -   I. An insect-specific venom produced in nature by a snake, a        wasp, etc. For example, see, Pang, et al., Gene, 116:165 (1992),        for disclosure of heterologous expression in plants of a gene        coding for a scorpion insectotoxic peptide.    -   J. An enzyme responsible for a hyperaccumulation of a        monoterpene, a sesquiterpene, a steroid, hydroxamic acid, a        phenylpropanoid derivative, or another non-protein molecule with        insecticidal activity.    -   K. An enzyme involved in the modification, including the        post-translational modification, of a biologically active        molecule; for example, a glycolytic enzyme, a proteolytic        enzyme, a lipolytic enzyme, a nuclease, a cyclase, a        transaminase, an esterase, a hydrolase, a phosphatase, a kinase,        a phosphorylase, a polymerase, an elastase, a chitinase, and a        glucanase, whether natural or synthetic. See, U.S. Pat. No.        5,955,653 which discloses the nucleotide sequence of a callase        gene. DNA molecules which contain chitinase-encoding sequences        can be obtained, for example, from the ATCC under Accession Nos.        39637 and 67152. See also, Kramer, et al., Insect Biochem.        Molec. Biol., 23:691 (1993), who teach the nucleotide sequence        of a cDNA encoding tobacco hornworm chitinase, and Kawalleck, et        al., Plant Molec. Biol., 21:673 (1993), who provide the        nucleotide sequence of the parsley ubi4-2 polyubiquitin gene,        U.S. Pat. Nos. 7,145,060, 7,087,810, and 6,563,020.    -   L. A molecule that stimulates signal transduction. For example,        see the disclosure by Botella, et al., Plant Molec. Biol.,        24:757 (1994), of nucleotide sequences for mung bean calmodulin        cDNA clones, and Griess, et al., Plant Physiol., 104:1467        (1994), who provide the nucleotide sequence of a maize        calmodulin cDNA clone.    -   M. A hydrophobic moment peptide. See, U.S. Pat. No. 5,580,852,        which discloses peptide derivatives of tachyplesin which inhibit        fungal plant pathogens, and U.S. Pat. No. 5,607,914 which        teaches synthetic antimicrobial peptides that confer disease        resistance.    -   N. A membrane permease, a channel former or a channel blocker.        For example, see the disclosure of Jaynes, et al., Plant Sci,        89:43 (1993), of heterologous expression of a cecropin-13 lytic        peptide analog to render transgenic tobacco plants resistant to        Pseudomonas solanacearum.    -   O. A viral-invasive protein or a complex toxin derived        therefrom. For example, the accumulation of viral coat proteins        in transformed plant cells imparts resistance to viral infection        and/or disease development effected by the virus from which the        coat protein gene is derived, as well as by related viruses.        See, Beachy, et al., Ann. Rev. Phytopathol., 28:451 (1990). Coat        protein-mediated resistance has been conferred upon transformed        plants against alfalfa mosaic virus, cucumber mosaic virus, and        tobacco mosaic virus.    -   P. An insect-specific antibody or an immunotoxin derived        therefrom. Thus, an antibody targeted to a critical metabolic        function in the insect gut would inactivate an affected enzyme,        killing the insect.    -   Q. A virus-specific antibody. See, for example, Tavladoraki, et        al., Nature, 366:469 (1993), who show that transgenic plants        expressing recombinant antibody genes are protected from virus        attack.    -   R. A developmental-arrestive protein produced in nature by a        pathogen or a parasite. Thus, fungal        endo-α-1,4-D-polygalacturonases facilitate fungal colonization        and plant nutrient release by solubilizing plant cell wall        homo-α-1,4-D-galacturonase. See, Lamb, et al., Bio/Technology,        10:1436 (1992). The cloning and characterization of a gene which        encodes a bean endopolygalacturonase-inhibiting protein is        described by Toubart, et al., Plant J., 2:367 (1992).    -   S. A developmental-arrestive protein produced in nature by a        plant. For example, Logemann, et al., Bio/Technology, 10:305        (1992), have shown that transgenic plants expressing the barley        ribosome-inactivating gene have an increased resistance to        fungal disease.    -   T. Genes involved in the Systemic Acquired Resistance (SAR)        Response and/or the pathogenesis-related genes. Briggs, S.,        Current Biology, 5(2) (1995); Pieterse & Van Loon, Curr. Opin.        Plant Bio., 7(4):456-64 (2004); and Somssich, Cell, 113(7):815-6        (2003).    -   U. Antifungal genes. See, Cornelissen and Melchers, Plant        Physiol., 101:709-712 (1993); Parijs, et al., Planta,        183:258-264 (1991); and Bushnell, et al., Can. J of Plant Path.,        20(2):137-149 (1998). See also, U.S. Pat. No. 6,875,907.    -   V. Detoxification genes, such as for fumonisin, beauvericin,        moniliformin, and zearalenone and their structurally-related        derivatives. See, U.S. Pat. No. 5,792,931.    -   W. Cystatin and cysteine proteinase inhibitors. See, U.S. Pat.        No. 7,205,453.    -   X. Defensin genes. See, U.S. Pat. Nos. 6,911,577, 7,855,327,        7855,328, 7,897,847, 7,910,806, 7,919,686, and 8,026,415.    -   Y. Genes conferring resistance to nematodes. See, U.S. Pat. Nos.        5,994,627 and 6,294,712; Urwin, et al., Planta, 204:472-479        (1998); Williamson, Curr Opin Plant Bio., 2(4):327-31 (1999).    -   Z. Genes that confer resistance to Phytophthora Root Rot, such        as the Rps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d, Rps 1-e, Rps        1-k, Rps 2, Rps 3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6, Rps        7, and other Rps genes. See, for example, Shoemaker, et al.,        Phytophthora Root Rot Resistance Gene Mapping in Soybean, Plant        Genome IV Conference, San Diego, Calif. (1995).    -   AA. Genes that confer resistance to Root Rot, such as described        by Williamson-Benavides, B.A., et al., “Identification of Root        Rot Resistance QTLs in Pea Using Fusarium solani f. sp.        pisi-Responsive Differentially Expressed Genes,” Front. Genet.        (Aug. 5, 2021).

    Any of the above-listed disease or pest resistance genes (A-AA) can    be introduced into the claimed field pea cultivar through a variety    of means including, but not limited to, transformation and crossing.-   2. Genes That Confer Resistance to an Herbicide, for Example:    -   A. An herbicide that inhibits the growing point or meristem,        such as an imidazolinone or a sulfonylurea. Exemplary genes in        this category code for mutant ALS and AHAS enzyme as described,        for example, by Lee, et al., EMBO J., 7:1241 (1988) and Mild, et        al., Theor. Appl. Genet., 80:449 (1990), respectively.    -   B. Glyphosate (resistance conferred by mutant        5-enolpyruvlshikimate-3-phosphate synthase (EPSPS) and aroA        genes, respectively) and other phosphono compounds, such as        glufosinate (phosphinothricin acetyl transferase (PAT) and        Streptomyces hygroscopicus PAT bar genes), pyridinoxy or phenoxy        proprionic acids, and cyclohexanediones (ACCase        inhibitor-encoding genes). See, for example, U.S. Pat. No.        4,940,835 which discloses the nucleotide sequence of a form of        EPSPS which can confer glyphosate resistance. U.S. Pat. No.        5,627,061 which describes genes encoding EPSPS enzymes. See        also, U.S. Pat. Nos. 6,566,587, 6,338,961, 6,248,876, 6,040,497,        5,804,425, 5,633,435, 5,145,783, 4,971,908, 5,312,910,        5,188,642, 4,940,835, 5,866,775, 6,225,114, 6,130,366,        5,310,667, 4,535,060, 4,769,061, 5,633,448, 5,510,471,        6,803,501, RE 36,449, RE 37,287, and 5,491,288, which are        incorporated herein by reference for this purpose. Glyphosate        resistance is also imparted to plants that express a gene that        encodes a glyphosate oxido-reductase enzyme, as described more        fully in U.S. Pat. Nos. 5,776,760 and 5,463,175, which are        incorporated herein by reference for this purpose. In addition,        glyphosate resistance can be imparted to plants by the over        expression of genes encoding glyphosate N-acetyltransferase.        See, for example, U.S. Pat. No. 7,462,481. A DNA molecule        encoding a mutant aroA gene can be obtained under ATCC Accession        No. 39256, and the nucleotide sequence of the mutant gene is        disclosed in U.S. Pat. No. 4,769,061. European Patent Appl. No.        0333033 and U.S. Pat. No. 4,975,374 disclose nucleotide        sequences of glutamine synthetase genes which confer resistance        to herbicides such as L-phosphinothricin. The nucleotide        sequence of a PAT gene is provided in European Patent No.        0242246 to Leemans, et al. DeGreef, et al., Bio/Technology,        7:61 (1989) describe the production of transgenic plants that        express chimeric bar genes coding for phosphinothricin acetyl        transferase activity. Exemplary of genes conferring resistance        to phenoxy proprionic acids and cyclohexones, such as sethoxydim        and haloxyfop are the Accl-S1, Accl-S2, and Acc2-S3 genes        described by Marshall, et al., Theor. Appl. Genet., 83:435        (1992).    -   C. An herbicide that inhibits photosynthesis, such as a triazine        (psbA and gs+ genes) and a benzonitrile (nitrilase gene).        Przibila, et al., Plant Cell, 3:169 (1991), describe the        transformation of Chlamydomonas with plasmids encoding mutant        psbA genes. Nucleotide sequences for nitrilase genes are        disclosed in U.S. Pat. No. 4,810,648 to Stalker and DNA        molecules containing these genes are available under ATCC        Accession Nos. 53435, 67441, and 67442. Cloning and expression        of DNA coding for a glutathione S-transferase is described by        Hayes, et al., Biochem. J., 285:173 (1992). Protoporphyrinogen        oxidase (PPO) is the target of the PPO-inhibitor class of        herbicides; a PPO-inhibitor resistant PPO gene was recently        identified in Amaranthus tuberculatus (Patzoldt et al., PNAS,        103(33):12329-2334, 2006). The herbicide methyl viologen        inhibits CO₂ assimilation. Foyer et al. (Plant Physiol.,        109:1047-1057, 1995) describe a plant overexpressing glutathione        reductase (GR) which is resistant to methyl viologen treatment.        Bromoxynil resistance by introducing a chimeric gene containing        the bxn gene (Science, 242(4877): 419-23, 1988).    -   D. Acetohydroxy acid synthase, which has been found to make        plants that express this enzyme resistant to multiple types of        herbicides, has been introduced into a variety of plants. See,        Hattori, et al., Mol. Gen. Genet., 246:419 (1995). Other genes        that confer tolerance to herbicides include a gene encoding a        chimeric protein of rat cytochrome P4507A1 and yeast        NADPH-cytochrome P450 oxidoreductase (Shiota, et al., Plant        Physiol., 106:17 (1994)); genes for glutathione reductase and        superoxide dismutase (Aono, et al., Plant Cell Physiol., 36:1687        (1995)); and genes for various phosphotransferases (Datta, et        al., Plant Mol. Biol., 20:619 (1992)).    -   E. Protoporphyrinogen oxidase (protox) is necessary for the        production of chlorophyll, which is necessary for all plant        survival. The protox enzyme serves as the target for a variety        of herbicidal compounds. These herbicides also inhibit growth of        all the different species of plants present, causing their total        destruction. The development of plants containing altered protox        activity which are resistant to these herbicides are described        in U.S. Pat. Nos. 6,288,306, 6,282,837, 5,767,373, and        6,084,155.

    Any of the above listed herbicide genes (A-E) can be introduced into    the claimed field pea cultivar through a variety of means including    but not limited to transformation and crossing.-   3. Genes That Confer or Contribute to a Value-Added Trait, such as:    -   A. Modified fatty acid metabolism, for example, by transforming        a plant with an antisense gene of stearyl-ACP desaturase to        increase stearic acid content of the plant. See, Knultzon, et        al., Proc. Natl. Acad. Sci. USA, 89:2625 (1992).    -   B. Decreased phytate content: 1) Introduction of a        phytase-encoding gene enhances breakdown of phytate, adding more        free phosphate to the transformed plant. For example, see, Van        Hartingsveldt, et al., Gene, 127:87 (1993), for a disclosure of        the nucleotide sequence of an Aspergillus niger phytase gene. 2)        Up-regulation of a gene that reduces phytate content. In maize,        this, for example, could be accomplished by cloning and then        re-introducing DNA associated with one or more of the alleles,        such as the LPA alleles, identified in maize mutants        characterized by low levels of phytic acid, such as in Raboy, et        al., Maydica, 35:383 (1990), and/or by altering inositol kinase        activity as in, for example, U.S. Pat. Nos. 7,425,442,        7,714,187, 6,197,561, 6,2191,224, 6,855,869, 6,391,348,        6,197,561, and 6,291,224; U.S. Publ. Nos. 2003/000901,        2003/0009011, and 2006/272046; and International Pub. Nos. WO        98/45448, and WO 01/04147.    -   C. Modified carbohydrate composition effected, for example, by        transforming plants with a gene coding for an enzyme that alters        the branching pattern of starch, or a gene altering thioredoxin,        such as NTR and/or TRX (See, U.S. Pat. No. 6,531,648, which is        incorporated by reference for this purpose), and/or a gamma zein        knock out or mutant, such as cs27 or TUSC27 or en27 (See, U.S.        Pat. Nos. 6,858,778, 7,741,533 and U.S. Publ. No. 2005/0160488,        which are incorporated by reference for this purpose). See,        Shiroza, et al., J. Bacteriol., 170:810 (1988) (nucleotide        sequence of Streptococcus mutans fructosyltransferase gene);        Steinmetz, et al., Mol. Gen. Genet., 200:220 (1985) (nucleotide        sequence of Bacillus subtilis levansucrase gene); Pen, et al.,        Bio/Technology, 10:292 (1992) (production of transgenic plants        that express Bacillus licheniformis α-amylase); Elliot, et al.,        PlantMolec. Biol., 21:515 (1993) (nucleotide sequences of tomato        invertase genes); Søgaard, et al., J. Biol. Chem.,        268:22480-22484 (1993) (site-directed mutagenesis of barley        α-amylase gene); Fisher, et al., Plant Physiol., 102:1045 (1993)        (maize endosperm starch branching enzyme II); International Pub.        No. WO 99/10498 (improved digestibility and/or starch extraction        through modification of UDP-D-xylose 4-epimerase, Fragile 1 and        2, Ref 1, HCHL, C4H); U.S. Pat. No. 6,232,529 (method of        producing high oil seed by modification of starch levels (AGP)).        The fatty acid modification genes mentioned above may also be        used to affect starch content and/or composition through the        interrelationship of the starch and oil pathways.    -   D. Elevated oleic acid via FAD-2 gene modification and/or        decreased linolenic acid via FAD-3 gene modification. See, U.S.        Pat. Nos. 5,952,544, 6,063,947, and 6,323,392. E. Altering        conjugated linolenic or linoleic acid content, such as in U.S.        Pat. No. 6,593,514. Altering LEC1, AGP, Dek1, Superal1, milps,        and various Ipa genes, such as Ipa1, Ipa3, hpt, or hggt. See,        for example, U.S. Pat. Nos. 7,122,658, 7,342,418, 6,232,529,        7,888,560, 6,423,886, 6,197,561, 6,825,397 and 7,157,621; U.S.        Publ. No. 2003/0079247; International Publ. No. WO 2003/011015;        and Rivera-Madrid, R., et al., Proc. Natl. Acad. Sci.,        92:5620-5624 (1995).    -   F. Altered antioxidant content or composition, such as        alteration of tocopherol or tocotrienols. See, for example, U.S.        Pat. Nos. 6,787,683, 7,154,029 and International Publ. No. WO        00/68393 (involving the manipulation of antioxidant levels        through alteration of a phytl prenyl transferase (ppt)); and        U.S. Pat. Nos. 7,154,029 and 7,622,658 (through alteration of a        homogentisate geranyl geranyl transferase (hggt)).    -   G. Altered essential seed amino acids. See, for example, U.S.        Pat. No. 6,127,600 (method of increasing accumulation of        essential amino acids in seeds); U.S. Pat. No. 6,080,913 (binary        methods of increasing accumulation of essential amino acids in        seeds); U.S. Pat. No. 5,990,389 and International Publ. No. WO        95/15392 (high lysine); U.S. Pat. No. 5,850,016 (alteration of        amino acid compositions in seeds); U.S. Pat. No. 5,885,802 (high        methionine); U.S. Pat. No. 5,885,801 and International Publ. No.        WO96/01905 (high threonine); U.S. Pat. No. 6,664,445, 7,022,895,        7,368,633, and 7,439,420 (plant amino acid biosynthetic        enzymes); U.S. Pat. No. 6,459,019 and U.S. Application No.        09/381.485 (increased lysine and threonine); U.S. Pat. No.        6,441,274 (plant tryptophan synthase beta subunit); U.S. Pat.        No. 6,346,403 (methionine metabolic enzymes); U.S. Pat. No.        5,939,599 (high sulfur); U.S. Pat. No. 5,912,414 (increased        methionine); U.S. Pat. No. 5,633,436 (increasing sulfur amino        acid content); U.S. Pat. No. 5,559,223 (synthetic storage        proteins with defined structure containing programmable levels        of essential amino acids for improvement of the nutritional        value of plants); U.S. Pat. No. 6,194,638 (hemicellulose); U.S.        Pat. No. 7,098,381 (UDPGdH); U.S. Pat. No. 6,194,638 (RGP); U.S.        Pat. Nos. 6,399,859, 6,930,225, 7,179,955, 6,803,498, 5,850,016,        and 7,053,282 (alteration of amino acid compositions in seeds);        WO 99/29882 (methods for altering amino acid content of        proteins); U.S. Application No. 09/297,418 (proteins with        enhanced levels of essential amino acids); WO 98/45458        (engineered seed protein having higher percentage of essential        amino acids); WO 01/79516; and U.S. Pat. Nos. 6,803,498,        6,930,225, 7,307,149, 7,524,933, 7,579,443, 7,838,632,        7,851,597, and 7,982,009 (maize cellulose synthases).-   4. Genes that Control Male Sterility: There are several methods of    conferring genetic male sterility available, such as multiple mutant    genes at separate locations within the genome that confer male    sterility, as disclosed in for example, Saxena, Kulbhushan, et al.,    “Male-sterility systems in pigeonpea and their role in enhancing    yield,” Plant Breeding. (April 2010). 129(2):125-134; and also see    U.S. Pat. Nos. 4,654,465 and 4,727,219 to Brar, et al., and    chromosomal translocations as described by Patterson in U.S. Pat.    Nos. 3,861,709 and 3,710,511. In addition to these methods,    Albertsen, et al., U.S. Pat. No. 5,432,068, describes a system of    nuclear male sterility which includes: identifying a gene which is    critical to male fertility; silencing this native gene which is    critical to male fertility; removing the native promoter from the    essential male fertility gene and replacing it with an inducible    promoter; inserting this genetically engineered gene back into the    plant; and thus creating a plant that is male sterile because the    inducible promoter is not “on” resulting in the male fertility gene    not being transcribed. Fertility is restored by inducing, or turning    “on,” the promoter, which in turn allows the gene that confers male    fertility to be transcribed.    -   A. Introduction of a deacetylase gene under the control of a        tapetum-specific promoter and with the application of the        chemical N—Ac—PPT. See, U.S. Pat. No. 6,384,304.    -   B. Introduction of various stamen-specific promoters. See, U.S.        Pat. Nos. 5,639,948 and 5,589,610.    -   C. Introduction of the barnase and the barstar genes. See, Paul,        et al., Plant Mol. Biol., 19:611-622 (1992).

    For additional examples of nuclear male and female sterility systems    and genes, see also, U.S. Pat. Nos. 5,859,341, 6,297,426, 5,478,369,    5,824,524, 5,850,014, and 6,265,640, all of which are hereby    incorporated by reference.-   5. Genes that Create a Site for Site Specific DNA Integration: This    includes the introduction of FRT sites that may be used in the    FLP/FRT system and/or Lox sites that may be used in the Cre/Loxp    system. See, for example, Lyznik, et al., Site-Specific    Recombination for Genetic Engineering in Plants, Plant Cell Rep,    21:925-932 (2003) and U.S. Pat. No. 6,187,994, which are hereby    incorporated by reference. Other systems that may be used include    the Gin recombinase of phage Mu (Maeser, et al. (1991); Vicki    Chandler, The Maize Handbook, Ch. 118 (Springer-Verlag 1994)); the    Pin recombinase of E. coli (Enomoto, et al. (1983)); and the R/RS    system of the pSRi plasmid (Araki, et al. (1992)).-   6. Genes that Affect Abiotic Stress Resistance: Genes that affect    abiotic stress resistance (including but not limited to flowering,    pod and seed development, enhancement of nitrogen utilization    efficiency, altered nitrogen responsiveness, drought resistance or    tolerance, cold resistance or tolerance, and salt resistance or    tolerance) and increased yield under stress. For example, see    Parmar, Nehanjali, et al, “Genetic engineering strategies for biotic    and abiotic stress tolerance and quality enhancement in    horticultural crops: a comprehensive review,” 3 Biotech. (2017 Aug).    7(4): 239; U.S. Pat. No. 6,653,535 where water use efficiency is    altered through alteration of malate; U.S. Pat. Nos. 5,892,009,    5,965,705, 5,929,305, 5,891,859, 6,417,428, 6,664,446, 6,706,866,    6,717,034, 6,801,104, 6,946,586, 7,238,860, 7,635,800, 7,135,616,    7,193,129, and 7,601,893; and International Publ. Nos. WO    2001/026459, WO 2001/035725, WO 2001/035727, WO 2001/036444, WO    2001/036597, WO 2001/036598, WO 2002/015675, and WO 2002/077185,    describing genes, including CBF genes and transcription factors    effective in mitigating the negative effects of freezing, high    salinity, and drought on plants, as well as conferring other    positive effects on plant phenotype; U.S. Publ. No. 2004/0148654,    where abscisic acid is altered in plants resulting in improved plant    phenotype, such as increased yield and/or increased tolerance to    abiotic stress; U.S. Pat. Nos. 6,992,237, 6,429,003, 7,049,115, and    7,262,038, where cytokinin expression is modified resulting in    plants with increased stress tolerance, such as drought tolerance,    and/or increased yield. See also, WO 02/02776, WO 2003/052063, JP    2002281975, U.S. Pat. No. 6,084,153, WO 01/64898, and U.S. Pat. Nos.    6,177,275 and 6,107,547 (enhancement of nitrogen utilization and    altered nitrogen responsiveness). For ethylene alteration, see, U.S.    Publ. Nos. 2004/0128719, 2003/0166197, and U.S. Application No.    09/856,834. For plant transcription factors or transcriptional    regulators of abiotic stress, see, i.e., U.S. Publ. Nos.    2004/0098764 or 2004/0078852.

Other genes and transcription factors that affect plant growth andagronomic traits, such as yield, flowering, plant growth, and/or plantstructure, can be introduced or introgressed into plants. See forexample, U.S. Pat. Nos. 6,140,085, and 6,265,637 (CO); U.S. Pat. No.6,670,526 (ESD4); U.S. Pat. Nos. 6,573,430 and 7,157,279 (TFL); U.S.Pat. No. 6,713,663 (FT); U.S. Pat. Nos. 6,794,560, 6,307,126 (GAI); U.S.Pat. No. 7,045,682 (VRN1); U.S. Pat. Nos. 6,949,694 and 7,253,274(VRN2); U.S. Pat. No. 6,887,708 (GI); U.S. Pat. No. 7,320,158 (FRI);U.S. Pat. No. 6,307,126 (GAI); U.S. Pat. Nos. 6,762,348 and 7,268,272(D8 and Rht); and U.S. Pat. Nos. 7,345,217, 7,511,190, 7,659,446, and7,825,296 (transcription factors).

Genetic Marker Profile Through SSR and First-Generation Progeny

In addition to phenotypic observations, a plant can also be identifiedby its genotype. The genotype of a plant can be characterized through agenetic marker profile which can identify plants of the same variety, ora related variety, or be used to determine or validate a pedigree.Genetic marker profiles can be obtained by techniques such asRestriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Amplified Fragment Length Polymorphisms(AFLPs), Simple Sequence Repeats (SSRs) (which are also referred to asMicrosatellites), and Single Nucleotide Polymorphisms (SNPs). See forexample, Gong, Yaming, et al., “Developing new SSR markers from ESTs ofpea (Pisum sativum L.),” J. Zhejiang Univ Sci B. (2010 September).11(9):702-707.

Particular markers used for these purposes are not limited to anyparticular set of markers, but are envisioned to include any type ofmarker and marker profile which provides a means of distinguishingvarieties. One method of comparison is to use only homozygous loci forfield pea cultivar 3997499.

Primers and PCR protocols for assaying these and other markers aredisclosed by for example, Jing, Runchun, et al., “Gene-Based SequenceDiversity Analysis of Field Pea (Pisum),” Genetics. (Dec. 1, 2007).177(4): 2263-2275. In addition to being used for identification of fieldpea variety 3997499, and plant parts and plant cells of field peavariety 3997499, the genetic profile may be used to identify a field peaplant produced through the use of field pea cultivar 3997499 or toverify a pedigree for progeny plants produced through the use of fieldpea cultivar 3997499. The genetic marker profile is also useful inbreeding and developing backcross conversions.

One embodiment comprises a field pea plant characterized by molecularand physiological data obtained from the sample of said varietydeposited with a Budapest Depository. Further provided by theembodiment(s) is a field pea plant formed by the combination of thedisclosed field pea plant or plant cell with another field pea plant orcell and comprising the homozygous alleles of the variety. “Cell” asused herein includes a plant cell, whether isolated, in tissue cultureor incorporated in a plant or plant part.

Means of performing genetic marker profiles using SSR polymorphisms arewell-known in the art. SSRs are genetic markers based on polymorphismsin repeated nucleotide sequences, such as microsatellites. A markersystem based on SSRs can be highly informative in linkage analysisrelative to other marker systems in that multiple alleles may be present(“linkage” refers to a phenomenon wherein alleles on the same chromosometend to segregate together more often than expected by chance if theirtransmission was independent). Another advantage of this type of markeris that, through use of flanking primers, detection of SSRs can beachieved, for example, by the polymerase chain reaction (PCR), therebyeliminating the need for labor-intensive Southern hybridization. The PCRdetection is done by use of two oligonucleotide primers flanking thepolymorphic segment of repetitive DNA. Repeated cycles of heatdenaturation of the DNA followed by annealing of the primers to theircomplementary sequences at low temperatures, and extension of theannealed primers with DNA polymerase, comprise the major part of themethodology.

Following amplification, markers can be scored by electrophoresis of theamplification products. Scoring of marker genotype is based on the sizeof the amplified fragment, which may be measured by the number of basepairs of the fragment. While variation in the primer used or inlaboratory procedures can affect the reported fragment size, relativevalues should remain constant regardless of the specific primer orlaboratory used. When comparing varieties, it is preferable if all SSRprofiles are performed in the same lab.

The SSR profile of field pea plant 3997499 can be used to identifyplants comprising field pea cultivar 3997499 as a parent, since suchplants will comprise the same homozygous alleles as field pea cultivar3997499. Because the field pea variety is essentially homozygous at allrelevant loci, most loci should have only one type of allele present. Incontrast, a genetic marker profile of an Fi progeny should be the sum ofthose parents, i.e., if one parent was homozygous for allele x at aparticular locus, and the other parent homozygous for allele y at thatlocus, then the Fi progeny will be xy (heterozygous) at that locus.Subsequent generations of progeny produced by selection and breeding areexpected to be of genotype x (homozygous), y (homozygous), or xy(heterozygous) for that locus position. When the Fi plant is selfed orsibbed for successive filial generations, the locus should be either xor y for that position.

In addition, plants and plant parts substantially benefiting from theuse of field pea cultivar 3997499 in their development, such as fieldpea cultivar 3997499 comprising a backcross conversion, transgene, orgenetic sterility factor, may be identified by having a molecular markerprofile with a high percent identity to field pea cultivar 3997499. Sucha percent identity might be 95%, 96%, 97%, 98%, 99%, 99.5%, or 99.9%identical to field pea cultivar 3997499. Percent identity refers to thecomparison of the homozygous alleles of two field pea varieties. Percentidentity or percent similarity is determined by comparing astatistically significant number of the homozygous alleles of twodeveloped varieties. For example, a percent identity of 90% betweenfield pea variety 1 and field pea variety 2 means that the two varietieshave the same allele at 90% of their loci.

The SSR profile of field pea cultivar 3997499 can also be used toidentify essentially derived varieties and other progeny varietiesdeveloped from the use of field pea cultivar 3997499, as well as cellsand other plant parts thereof. Such plants may be developed using themarkers, for example, by Teshome, Abel, et al., “Assessment of geneticdiversity in Ethiopian field pea (Pisum sativum L.) accessions withnewly developed EST-SSR markers,” BMC Genetics. (2015). 16: Articlenumber: 102.

Progeny plants and plant parts produced using field pea cultivar 3997499may be identified by having a molecular marker profile of at least 25%,30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 78%, 79%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99%, or 99.5% genetic contribution from fieldpea variety, as measured by either percent identity or percentsimilarity. Such progeny may be further characterized as being within apedigree distance of field pea cultivar 3997499, such as within 1, 2, 3,4, or 5 or less cross-pollinations to a field pea plant other than fieldpea cultivar 3997499 or a plant that has field pea cultivar 3997499 as aprogenitor. Unique molecular profiles may be identified with othermolecular tools such as SNPs and RFLPs.

While determining the SSR genetic marker profile of the plants describedsupra, several unique SSR profiles may also be identified which did notappear in either parent of such plant. Such unique SSR profiles mayarise during the breeding process from recombination or mutation. Acombination of several unique alleles provides a means of identifying aplant variety, an Fi progeny produced from such variety, and progenyproduced from such variety. Tissue Culture

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of field pea andregeneration of plants therefrom is well-known and widely published. Forexample, see Rubluo, A., et al., “Plant Regeneration from Pea LeafletsCultured in vitro and Genetic Stability of Regenerants,” Journal ofPlant Physiology. (December 1984). 117(2): 119-130; Ghanem, S.A., etal., “In vitro studies on Pea (Pisum sativum L.): I. Callus formation,regeneration and rooting,” Giornale botanico italiano. (1996). 130(4-6):695-705; and Bobkov, Sergey, “Obtaining Calli and Regenerated Plants inAnther Cultures of Pea,” Czech J. Genet. Plant Breed. (2014). 50(2):123-129.

Journal homepage. Thus, another aspect or embodiment is to provide cellswhich upon growth and differentiation produce field pea plants havingthe physiological and morphological characteristics of field peacultivar 3997499.

Regeneration refers to the development of a plant from tissue culture.The term “tissue culture” indicates a composition comprising isolatedcells of the same or a different type or a collection of such cellsorganized into parts of a plant. Exemplary types of tissue cultures areprotoplasts, calli, plant clumps, and plant cells that can generatetissue culture that are intact in plants or parts of plants, such asembryos, pollen, flowers, seeds, pods, petioles, leaves, stems, roots,root tips, anthers, pistils, and the like. Means for preparing andmaintaining plant tissue culture are well known in the art. By way ofexample, a tissue culture comprising organs has been used to produceregenerated plants. Hussain, Altaf, et al., “Plant Tissue Culture:Current Status and Opportunities,” in Recent Advances in Plant in vitroCulture. Published October 17th 2012. Additionally, U.S. Pat. Nos.5,959,185, 5,973,234, and 5,977,445 describe certain techniques, thedisclosures of which are incorporated herein by reference.

Plant Products

The seed of field pea cultivar 3997499, the plant produced from theseed, the hybrid field pea plant produced from the crossing of thevariety with any other field pea plant, hybrid seed, and various partsof the hybrid field pea plant can be utilized for plant products.Industrial uses include but are not limiting to, human food, livestockfeed, and as a raw material in industry.

As used herein the term plant product will be understood to mean theproduct derived from or produced by a plant of field pea cultivar3997499, for example, the tissues or structures of the plant of fieldpea cultivar 3997499 such as the flower, fruit, seed, leaves, stemsetc., produced by the plant. Further, the field pea seeds produced fromor derived from field pea cultivar 3997499 can be crushed, or acomponent of the field pea seeds can be extracted, in order to comprisea plant product, such as protein concentrate, protein isolate, meal,flour, and for a food or feed product. In other embodiments, a processedproduct includes, but is not limited to: dehydrated, cut, sliced,ground, pureed, dried, baked, fried, canned, jarred, washed, brined,packaged, refrigerated, frozen and/or heated pods, and/or seeds of thepea plants of the invention, or any other part thereof. In furtherembodiments, a processed product includes a sugar or other carbohydrate,fiber, protein and/or aromatic compound that is extracted, purified orisolated from pea plants disclosed herein. In embodiments, the processedproduct includes washed and packaged pods and/or seeds (or partsthereof) of the field pea cultivar 3997499 for example, in a canned orfrozen form. In other embodiments, the processed product is a whole podthat has been dehydrated and/or baked.

The field pea may also be used an oilseed crop. See for example,Villalobos Solis, M.I., et al., “Fatty acid profiling of the seed oilsof some varieties of field peas (Pisum sativum) by RP-LC/ESI-MS/MS:towards the development of an oilseed pea Changes in fatty acidcomposition for improved oxidative stability and nutrition areconstantly sought after,” Food Chem. (2013). 139(1-4): 986-989. Thus,potential industrial uses of field pea oil, which is subjected tofurther processing, include ingredients for paints, plastics, fibers,detergents, cosmetics, lubricants, and biodiesel fuel. Pea oil may besplit, inter-esterified, sulfurized, epoxidized, polymerized,ethoxylated, or cleaved. Designing and producing field pea oilderivatives with improved functionality and improved oleochemistry is arapidly growing field. The typical mixture of triglycerides is usuallysplit and separated into pure fatty acids, which are then combined withpetroleum-derived alcohols or acids, nitrogen, sulfonates, chlorine, orwith fatty alcohols derived from fats and oils to produce the desiredtype of oil or fat.

Field pea cultivar 3997499 can be used to produce field pea oil. Toproduce field pea oil, the seeds harvested from field pea cultivar3997499 may be for example, cracked, adjusted for moisture content,rolled into flakes and the oil is solvent-extracted from the flakes withcommercial hexane. The oil is then refined, blended for differentapplications, and sometimes hydrogenated. The resulting oils, bothliquid and partially hydrogenated, may be used domestically andexported, sold as oil or are used in a wide variety of processed foods.

Field peas are also used as a food source for both animals and humans.Field peas are widely used as a source of protein for animal feed.

Field pea cultivar 3997499 can be used to produce meal. Bansal, P., etal., “Dry Pea: Production Driven by Demand for Animal Feed,” (2019). InD. K. Navarro & V. Rawal (Eds.), The Global Economy of Pulses, 1st ed.(pp. 99-106). Rome: FAO. Field pea meal produced from field pea cultivar3997499 can also be used to field pea protein concentrate and field peaprotein isolate.

In addition, field pea cultivar 3997499 can be used to produce peaflour. Beitane, I., et al., “Dietary micronutrient content in pea (PisumSativum L.) and buckwheat (Fagopyrum Esculentum M.) flour,” (2017). InE. Straumite (Ed.), 11th Baltic Conference on Food Science andTechnology ‘Food science and technology in a changing world (Vol. 700,pp. 56-60); and Krumina-Zemture, G., et al., “Amino acid and dietaryfibre content of pea and buckwheat flours,” (2016). In Research forRural Development, 18-20 May 2016, (pp. 84-90). Jelgava: LatviaUniversity of Agriculture, where pea flour was observed to be a goodsource (in comparison to wheat) of vitamins Bi and B₂.

Additionally, field pea cultivar 3997499 can be used to produce varioustypes of “fillers” in food products. Examples of food productscontaining pea derived products are protein powder, meat-freeburgers/minced meat/sausages, dairy alternative drinks, yogurtalternatives, vegan cheese and puffs, and protein bars. See, Rasskazova,I., et al., “Field pea Pisum sativum L. as a perspective ingredient forvegan foods; a review,” Food Science. (2020). 35: 125-131. Thus, thefield peas produced by field pea cultivar 3997499 can be processed toproduce a texture and appearance similar to many other foods.

High Protein Content

For consumption, field pea cultivar 3997499 can be used to produceedible protein ingredients which offer a healthier, less expensivereplacement for animal protein in meats, as well as in dairy-typeproducts. Field peas are approximately 21.2-32.9% protein, 36.9%-49%starch, and 14-26% dietary fiber, by dry weight. See, Geerts, M.E.J., etal, “Mildly refined fractions of yellow peas show rich behaviour inthickened oil-in-water emulsions,” Innovative Food Science and EmergingTechnologies. (2017). 41: 251-258; Lan, Y., et al., “Soliddispersion-based spray-drying improves solubility and mitigates beanyflavour of pea protein isolate,” Food Chemistry. (2019). 278: 665-673;Pietrasik, Z., et al., “Utilization of pea starch and fibre fractionsfor replacement of wheat crumb in beef burgers,” Meat Science. (October2019). 161: 107974. Peas being a legume crop have two aspects thatdistinguish them from most other food crops. Firstly, they are rich withmacro and micronutrients: being a good source of protein (rich withessential amino acids as tryptophan and lysine), slowly digestiblecarbohydrates, B group vitamins, minerals, dietary fiber (soluble andinsoluble), phytosterols, and α-linolenic, acid. They also provide someamounts of squalene, tocopherols, polyphenols and triterpenic acids.

Compared to soybean or other proteins derived from plants, pea proteinis associated with being more digestible and having relatively lessallergenic responses and negative health controversies. See for example,Owasu-Ansah, Y.J., et al., “Pea proteins: a review of chemistry,technology of production, and utilization,” Food Reviews International.1991. 7(1): 13-134 and Allred, C.D., et al., “Soy processing influencesgrowth of estrogen-dependent breast cancer tumors,” Carcinogenesis.2004. 25(9): 1649-1657. Thus, another embodiment is to create highprotein varieties using one or more of field pea cultivar 3997499.

Agricultural Treatment Agents

A plant, or its environment, can be contacted with a wide variety of“agriculture treatment agents.” As used herein, an “agriculturetreatment agent”, or “treatment agent”, or “agent” can refer to anyexogenously provided compound that can be brought into contact with aplant tissue (e.g., a seed) or its environment that affects a plant’sgrowth, development and/or performance, including agents that affectother organisms in the plant’s environment when those effectssubsequently alter a plant’s performance, growth, and/or development(e.g., an insecticide that kills plant pathogens in the plant’senvironment, thereby improving the ability of the plant to tolerate theinsect’s presence). Agriculture treatment agents also include a broadrange of chemicals and/or biological substances that are applied toseeds, in which case they are commonly referred to as seed treatmentsand/or seed dressings. Seed treatments are commonly applied as either adry formulation or a wet slurry or liquid formulation prior to plantingand, as used herein, generally include any agriculture treatment agentincluding growth regulators, micronutrients, nitrogen-fixing microbes,and/or inoculants. Agriculture treatment agents include pesticides(e.g., fungicides, insecticides, bactericides, etc.) hormones (abscisicacids, auxins, cytokinins, gibberellins, etc.) herbicides (e.g.,glyphosate, atrazine, 2,4-D, dicamba, etc.), nutrients (e.g., a plantfertilizer), and/or a broad range of biological agents, for example aseed treatment inoculant comprising a microbe that improves cropperformance, i.e., by promoting germination and/or root development. Incertain embodiments, the agriculture treatment agent actsextracellularly within the plant tissue, such as interacting withreceptors on the outer cell surface. In some embodiments, theagriculture treatment agent enters cells within the plant tissue.

In certain embodiments, the agriculture treatment agent remains on thesurface of the plant and/or the soil near the plant. In certainembodiments, the agriculture treatment agent is contained within aliquid. Such liquids include, but are not limited to, solutions,suspensions, emulsions, and colloidal dispersions. In some embodiments,liquids described herein will be of an aqueous nature. However, invarious embodiments, such aqueous liquids that comprise water can alsocomprise water insoluble components, can comprise an insoluble componentthat is made soluble in water by addition of a surfactant, or cancomprise any combination of soluble components and surfactants. Incertain embodiments, the application of the agriculture treatment agentis controlled by encapsulating the agent within a coating, or capsulee.g., microencapsulation). In certain embodiments, the agriculturetreatment agent comprises a nanoparticle and/or the application of theagriculture treatment agent comprises the use of nanotechnology.

While a number of exemplary aspects and embodiments have been discussedabove, those of skill in the art will recognize certain modifications,permutations, additions and sub-combinations thereof. It is thereforeintended that the following appended claims and claims hereafterintroduced are interpreted to include all such modifications,permutations, additions, and sub-combinations as are within their truespirit and scope.

One embodiment may be embodied in other specific forms without departingfrom its spirit or essential characteristics. The described embodimentsare to be considered in all respects only as illustrative and notrestrictive. All changes which come within the meaning and range ofequivalency of the claims are to be embraced within their scope.

Various embodiments, include components, methods, processes, systemsand/or apparatus substantially as depicted and described herein,including various embodiments, sub-combinations, and subsets thereof.Those of skill in the art will understand how to make and use anembodiment(s) after understanding the present disclosure.

The foregoing discussion of the embodiments has been presented forpurposes of illustration and description. The foregoing is not intendedto limit the embodiments to the form or forms disclosed herein. In theforegoing Detailed Description for example, various features of theembodiments are grouped together in one or more embodiments for thepurpose of streamlining the disclosure. This method of disclosure is notto be interpreted as reflecting an intention that the embodiment(s)requires more features than are expressly recited in each claim. Rather,as the following claims reflect, inventive aspects lie in less than allfeatures of a single foregoing disclosed embodiment. Thus, the followingclaims are hereby incorporated into this Detailed Description.

Moreover, though the description of the embodiments has includeddescription of one or more embodiments and certain variations andmodifications, other variations and modifications are within the scopeof the embodiments (e.g., as may be within the skill and knowledge ofthose in the art, after understanding the present disclosure). It isintended to obtain rights which include alternative embodiments to theextent permitted, including alternate, interchangeable and/or equivalentstructures, functions, ranges or acts to those claimed, whether or notsuch alternate, interchangeable and/or equivalent structures, functions,ranges or acts are disclosed herein, and without intending to publiclydedicate any patentable subject matter.

The terms “comprising,” “having,” “including,” and “containing” are tobe construed as open-ended terms (i.e., meaning “including, but notlimited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. Forexample, if the range 10-15 is disclosed, then 11, 12, 13, and 14 arealso disclosed. All methods described herein can be performed in anysuitable order unless otherwise indicated herein or otherwise clearlycontradicted by context. The use of any and all examples, or exemplarylanguage (e.g., “such as”) provided herein, is intended merely to betterilluminate the embodiments and does not pose a limitation on the scopeof the embodiments unless otherwise claimed.

DEPOSIT INFORMATION

A deposit of the Benson Hill Seeds, Inc. proprietary field pea cultivar3997499 disclosed above and recited in the appended claims is maintainedby Benson Hill Seeds, Inc. A deposit will be made withProvasoli-Guillard National Center for Marine Algae and Microbiota,Bigelow Laboratory for Ocean Sciences (NCMA). Access to this depositwill be available during the pendency of this application to personsdetermined by the Commissioner of Patents and Trademarks to be entitledthereto under 37 C.F.R. 1.14 and 35 U.S.C. §122. Upon allowance of anyclaims in this application, all restrictions on the availability to thepublic of the variety will be irrevocably removed by affording access toa deposit of at least 625 seeds of the same variety with NCMA. Thedeposit will be maintained in the depository for a period of 30 years,or 5 years after the last request, or for the effective life of thepatent, whichever is longer, and will be replaced if necessary, duringthat period.

What is claimed is:
 1. A plant or a seed of field pea cultivar 3997499,wherein a representative sample of seed of said cultivar is depositedunder NCMA No.______.
 2. A field pea plant, or a part thereof, of theplant or seed of claim 1, wherein the plant or plant part comprises atleast one cell of field cultivar
 3997499. 3. A tissue culture comprisingat least one cell or protoplast of the plant or plant part of claim 2.4. A method for producing a field pea seed, wherein the method comprisesfertilizing a field pea plant and harvesting the resultant field peaseed, wherein the field pea plant is the field pea plant of claim
 1. 5.A field pea seed produced by the method of claim 4, wherein theresulting field pea seed is an F₁ offspring or the product ofself-fertilization.
 6. A field pea plant, or a part thereof, produced bygrowing the seed of claim
 5. 7. A method of producing a plant derivedfrom field pea cultivar 3997499 comprising an added desired trait,wherein the method comprises introducing at least one mutation in anucleic acid sequence of the field pea plant, or plant part thereof, orseed of claim 1, wherein the mutation confers the desired trait to atleast one cell of field pea cultivar
 3997499. 8. A field pea plantproduced by the method of claim 7, wherein the plant comprises thedesired trait.
 9. A method of producing a plant derived from field peacultivar 3997499, wherein a representative sample of seed of saidcultivar is deposited under NCMA No. ______. comprising an added desiredtrait, wherein the method comprises introducing at least one nucleicacid sequence conferring the desired trait to said plant.
 10. A fieldpea plant produced by the method of claim 9, wherein the plant comprisesthe desired trait.
 11. The field pea plant of claim 10, wherein thedesired trait is selected from the group comprising male sterility,herbicide tolerance, pest tolerance, disease tolerance, modified fattyacid metabolism, modified carbohydrate metabolism, modified seed yield,modified seed oil, modified seed protein, modified shattering, modifiediron-deficiency chlorosis, modified water use efficiency, and/orcombinations thereof.
 12. A method of producing a plant productcomprising collecting a plant product from the plant of claim 1, orplant part thereof.
 13. A field pea plant product produced by the methodof claim 12, wherein the plant product is produced from a plant havingat least one cell of field pea cultivar
 3997499. 14. A method ofintroducing a desired trait into field pea cultivar 3997499, wherein themethod comprises: (a) crossing a 3997499 plant, wherein a sample of seedis deposited under NCMA No. ______, with a plant of another field peacultivar having a desired trait to produce progeny plants; (b) selectingone or more progeny plants that have the desired trait to produceselected progeny plants; (c) crossing the selected progeny plants withthe 3997499 plant to produce backcross progeny plants; (d) selecting forbackcross progeny plants that have the desired trait; and (e) repeatingsteps (c) and (d) a sufficient number of times in succession to produceselected second or higher backcross progeny plants that comprise thedesired trait and essentially all of the physiological and morphologicalcharacteristics of field pea cultivar
 3997499. 15. A field pea plantproduced by the method of claim 14 wherein the plant has the desiredtrait.
 16. A method for developing a field pea plant, comprisingapplying plant breeding techniques to the plant of claim 1, or plantpart thereof, comprising crossing, recurrent selection, mutationbreeding, wherein said mutation breeding selects for a mutation that isspontaneous or artificially induced, backcrossing, genomic selection,pedigree breeding, marker enhanced selection, haploid/double haploidproduction, speed breeding, apomixis, or transformation, whereinapplication of said techniques results in development of a new field peaplant.
 17. A method of introducing a mutation into the genome of fieldpea cultivar 3997499, said method comprising mutagenesis of the plant ofclaim 1, or a part thereof.
 18. A method of editing the genome of fieldpea cultivar 3997499, said method comprising editing the genome of theplant, of claim 1, or plant part thereof, wherein said method isselected from the group comprising zinc finger nucleases, transcriptionactivator-like effector nucleases (TALENs), engineered homingendonucleases/meganucleases, and the clustered regularly interspacedshort palindromic repeat (CRISPR)-associated protein9 (Cas9) system. 19.A field pea plant produced by the method of claim
 18. 20. The seed, orpart thereof, of claim 1, wherein the seed further comprises at leastone seed treatment.